Methods of using thiazolidine derivatives to treat cancer or inflammation

ABSTRACT

Methods of using thiazolidine derivatives of formula (I) to treat cancer, inflammation, or other disorders related to the activities of protein phosphatases PTPN12 or PTPN2 in a mammal are disclosed. Pharmaceutical compositions containing such derivatives are disclosed.

FIELD OF THE INVENTION

This invention is directed to methods of using thiazolidine derivativesto treat cancer or inflammation in a mammal.

BACKGROUND OF THE INVENTION

Protein tyrosine phosphorylation is an important mechanism fortransmitting extracellular stimuli in biochemical and cellular eventssuch as cell attachment, mitogenesis, differentiation and migration (seee.g., Li, Liwu et al., Seminars in Immunology (2000), Vol. 12, pp.75-84, and Neel, B. G. et al., Current Opinion in Cell Biology (1997),Vol. 9, pp. 193-204). Kinases and phosphatases are enzymes that helpregulate many cellular activities, particularly signaling from the cellmembrane to the nucleus to initiate the cell's entrance into the cellcycle and to control other functions:

Phosphorylation is important in signal transduction mediated byreceptors via extracellular biological signals such as growth factors orhormones. For example, many oncogenes are kinases or phosphatases, i.e.enzymes that catalyze protein phosphorylation or dephosphorylationreactions or are specifically regulated by phosphorylation. In addition,a kinase or phosphatase can have its activity regulated by one or moredistinct kinase or phosphatases, resulting in specific signalingcascades.

All protein tyrosine phosphatases (PTPs) have a conserved catalyticdomain characterized by a signature sequence (I/V)HCXXGXX(S/T).Biochemical and kinetic studies have demonstrated that the cysteineresidue found in this signature sequence is essential for catalyticactivity of PTPs since this mutation of this cysteine completelyabolishes PTP activity. See, Flint, A. J., et al., Proc. Natl. Acad.Sci. U.S.A. 94 (1997), pp. 1680-1685.

DESCRIPTION OF THE RELATED ART

PCT Published Patent Application, WO 99/61467 (McGill University),describes agents that interfere with the binding of PTPN12 (PTP-PEST) todomains of signalling proteins as inhibitors of cell migration and/or offocal adhesion.

PCT Published Patent Application, WO 00/36111 (McGill University)describes methods of utilizing PTPN2 (TC-PTP) for screening.

U.S. Pat. No. 5,726,027 by Olefsky, Jerrold M. describes a screeningmethod for identifying compositions which affect the binding of proteintyrosine phosphatase 1B (PTP1B)

U.S. Pat. No. 6,262,044 (Novo Nordisk) describes certain proteintyrosine phosphatase inhibitors and provides a detailed description ofthe discovery of protein tyrosine phosphatases and theirpathophysiological roles.

Gorishnil, V. Ya. et al, Farm. Zh. (Kiev) (2001), Vol. 2, pp. 6467, andGorishnyi, V. Ya. et al, Farm. Zh. (Kiev) (1995), Vol. 4, pp. 50-53,discloses 4-oxo-2-thioxothiazolidine derivatives useful in treatinginflammation.

PCT Published Patent Application WO 00/76988 (Warner-Lambert) discloses4-oxo-2-thioxothiazolidine derivatives useful as amyloid aggregationinhibitors and in imaging amyloid deposits.

European Patent Specification 0047109 (Ono Pharmaceuticals) discloses4-oxo-2-thioxothiazolidine derivatives useful in inhibiting aldosereductase.

SUMMARY OF THE INVENTION

This invention is directed to the use of certain thiazolidinederivatives in treating cancer and inflammation in a mammal.

Accordingly, one aspect of this invention provides a method of treatingcancer in a mammal, which method comprises administering to the mammalin need thereof a therapeutically effective amount of a compound offormula (I):

wherein:

each p is independently 1 to 5;

A is a linker of four atoms and is selected from the group consisting ofan optionally substituted straight or branched alkylene chain of fourcarbons, an optionally substituted straight or branched alkenylene chainof four carbons, —R⁸—C(R²)—N(R⁵)—, —R⁸—N(R⁵)—C(R²)—, —R⁸—O—C(R²)—,R⁷—O—C(R²)—R⁷—, —R⁸—O—R⁷—, —R⁷—C(R²)—N(R⁵)—S(O)_(t)— (where t is 0 to2), —R⁸—N(R⁵)—R⁷, —R⁸—S(O)_(t)—R⁷— (where t is 0 to 2), —R⁹—N(R⁵)—,—R⁹—O—, and —R⁹—C(R²)—;

R¹ and R² are each independently ═O or ═S;

each R³ and R⁶ are independently selected from the group consisting ofhydrogen, alkyl, alkenyl, aryl, aralkyl, aralkenyl, cycloalkyl,cycloalkylalkyl, halo, haloalkyl, haloalkoxy, nitro, cyano, —N═N—O—R¹¹,—OR¹⁰, —C(O)OR¹⁰, —C(O)N(R¹⁰)₂, —N(R¹⁰)₂, —N(R¹⁰)C(O)R¹⁰,—N(R¹⁰)C(O)OR¹¹, —S(O)_(t)R¹⁰ (where t is 0 to 2), —S(O)_(t)N(R¹⁰)₂(where t is 0 to 2), —S(O)_(t)NH—R¹⁴, heterocyclyl andheterocyclylalkyl;

R⁴ is hydrogen, alkyl, aralkyl, aryl, haloalkyl, cycloalkyl,cycloalkylalkyl, heterocyclyl or heterocyclylalkyl;

each R⁵ is independently hydrogen, alkyl, aralkyl, or aryl;

each R⁷ is an optionally substituted alkylene chain of one carbon;

each R⁸ is an optionally substituted straight or branched alkylene oralkenylene chain of two carbons;

each R⁹ is an optionally substituted alkylene chain of three carbons;

each R¹⁰ is hydrogen, alkyl, alkenyl, cycloalkyl, cycloalkylalkyl,aralkyl or aryl; and

R¹¹ is hydrogen, alkyl or aralkyl;

R¹⁴ is a thiazole;

as a single stereoisomer, a mixture of stereoisomers, or as a racemicmixture of stereoisomers; or as a solvate or polymorph; or as apharmaceutically acceptable salt thereof.

In another aspect, this invention provides a method of treatinginflammation in a mammal, which method comprises administering to themammal in need thereof a therapeutically effective amount of a compoundof formula (Ia):

wherein:

each p is independently 1 to 5;

A is linker of four atoms and is selected from the group consisting ofan optionally substituted straight or branched alkylene chain of fourcarbons, an optionally substituted straight or branched alkenylene chainof four carbons, —R^(8a)—C(R^(2a))—N(R^(5a))—,—R^(8a)—N(R^(5a))—C(R^(2a))—, —R^(8a)—O—C(R^(2a))—,R^(7a)—O—C(R^(2a))—R^(7a), R^(8a)—O—R^(7a)—,—R^(7a)—C(R^(2a))—N(R^(5a))—S(O)_(t)— (where t is 0 to 2),—R^(8a)—N(R^(5a))—R^(7a), —R^(8a)—S(O)_(t)—R^(7a)— (where t is 0 to 2),—R^(9a)—N(R^(5a))—, —R^(9a)—O—, and —R^(9a)—C(R^(2a))—;

R^(1a) and R^(2a) are each independently ═O or ═S;

R^(3a) is alkyl, alkenyl, aryl, aralkyl, aralkenyl, cycloalkyl,cycloalkylalkyl, chloro, iodo, bromo, haloalkyl, haloalkoxy, nitro,cyano, —N═N—O—R^(11a), —OR^(10a), —C(O)OR^(10a), —C(O)N(R^(10a))₂,—N(R^(12a))₂, —N(R^(10a))C(O)R^(10a), —N(R^(10a))C(O)OR^(11a),—S(O)_(t)R^(10a) (where t is 0 to 2), S(O)_(t)N(R^(10a))₂ (where t is 0to 2), or heterocyclylalkyl;

R^(4a) is hydrogen, alkyl, aralkyl, aryl, haloalkyl, cycloalkyl,cycloalkylalkyl, heterocyclyl or heterocyclylalkyl;

R^(5a) is hydrogen, alkyl, aralkyl, or aryl;

R^(6a) is alkyl, alkenyl, aryl, aralkyl, aralkenyl, cycloalkyl,cycloalkylalkyl, halo, haloalkyl, haloalkoxy, nitro, cyano,—N═N—O—R^(11a), —C(O)N(R^(10a))₂, N(R^(10a))₂,—N(R^(10a))C(O)R^(10a)—N(R^(10a))C(O)OR^(11a), —S(O)_(t)R^(10a) (where tis 0 to 2), —S(O)_(t)N(R^(10a))₂ (where t is 0 to 2),—S(O)_(t)NH—R^(14a), heterocyclyl or heterocyclylalkyl;

each R^(7a) is an optionally substituted alkylene chain of one carbon;

each R^(8a) is an optionally substituted straight or branched alkyleneor alkenylene chain of two carbons;

each R^(9a) is an optionally substituted alkylene chain of threecarbons;

each R^(10a) is hydrogen, alkyl, alkenyl, cycloalkyl, cycloalkylalkyl,aralkyl or aryl;

R^(11a) is hydrogen, alkyl or aralkyl; and

R^(12a) is hydrogen, aryl or aralkyl;

R^(14a) is a thiazole;

as a single stereoisomer, a mixture of stereoisomers, or as a racemicmixture of stereoisomers; or as a solvate or polymorph; or as apharmaceutically acceptable salt thereof.

In another aspect, this invention provides a method of treating a mammalhaving a disorder or condition associated with hyperproliferation andtissue remodelling or repair, wherein said method comprisesadministering to the mammal having the disorder or condition atherapeutically effective amount of a compound of formula (I):

wherein:

each p is independently 1 to 5;

A is a linker of four atoms and is selected from the group consisting ofan optionally substituted straight or branched alkylene chain of fourcarbons, an optionally substituted straight or branched alkenylene chainof four carbons, —R⁸—C(R²)—N(R⁵)—, —R⁸—N(R⁵)—C(R²)—, —R⁸—O—C(R²)—,R⁷—O—C(R²)—R⁷—, —R⁸—R⁷—, —R⁷—C(R²)—N(R⁵)—S(O)_(t)— (where t is 0 to 2),—R⁸—N(R⁵)—R⁷, —R⁸—S(O)_(t)—R⁷— (where t is 0 to 2), —R⁹—N(R⁵)—, —R⁹—O—,and —R⁹—C(R²)—;

R¹ and R² are each independently ═O or ═S;

each R³ and R⁶ are independently selected from the group consisting ofhydrogen, alkyl, alkenyl, aryl, aralkyl, aralkenyl, cycloalkyl,cycloalkylalkyl, halo, haloalkyl, haloalkoxy, nitro, cyano, —N═N—O—R¹¹,—OR¹⁰, —C(O)OR¹⁰, —C(O)N(R¹⁰)₂, —N(R¹⁰)₂, —N(R¹⁰)C(O)R¹⁰,—N(R¹⁰)C(O)OR¹¹, —S(O)_(t)R¹⁰ (where t is 0 to 2), —S(O)_(t)N(R¹⁰)₂(where t is 0 to 2), heterocyclyl and heterocyclylalkyl;

R⁴ is hydrogen, alkyl, aralkyl, aryl, haloalkyl, cycloalkyl,cycloalkylalkyl, heterocyclyl or heterocyclylalkyl;

each R⁵ is independently hydrogen, alkyl, aralkyl, or aryl;

each R⁷ is an optionally substituted alkylene chain of one carbon;

each R⁸ is an optionally substituted straight or branched alkylene oralkenylene chain of two carbons;

each R⁹ is an optionally substituted alkylene chain of three carbons;

each R¹⁰ is hydrogen, alkyl, alkenyl, cycloalkyl, cycloalkylalkyl,aralkyl or aryl; and

R¹¹ is hydrogen, alkyl or aralkyl;

as a single stereoisomer, a mixture of stereoisomers, or as a racemicmixture of stereoisomers; or as a solvate or polymorph; or as apharmaceutically acceptable salt thereof.

In another aspect, this invention provides a method of treating amammalian cell with a compound of formula (I):

wherein:

each p is independently 1 to 5;

A is a linker of four atoms and is selected from the group consisting ofan optionally substituted straight or branched alkylene chain of fourcarbons, an optionally substituted straight or branched alkenylene chainof four carbons, —R⁸—C(R²)—N(R⁵)—, —R⁸—N(R⁵)—C(R²)—, —R⁸—O—C(R²)—,—R⁷—O—C(R²)—R⁷—, —R⁸—O—R⁷—, R⁷—C(R²)—N(R⁵)—S(O)_(t)— (where t is 0 to2), —R⁸—N(R⁵)—R⁷, —R⁸—S(O)_(t)—R⁷— (where t is 0 to 2), —R⁹—N(R⁵)—,—R⁹—O—, and —R⁹—C(R²)—;

R¹ and R² are each independently ═O or ═S;

each R³ and R⁶ are independently selected from the group consisting ofhydrogen, alkyl, alkenyl, aryl, aralkyl, aralkenyl, cycloalkyl,cycloalkylalkyl, halo, haloalkyl, haloalkoxy, nitro, cyano, —N═N—O—R¹¹,—OR¹⁰, —C(O)OR¹⁰, —C(O)N(R¹⁰)₂, —N(R¹⁰)₂, —N(R¹⁰)C(O)R¹⁰,—N(R¹⁰)C(O)OR¹¹, —S(O)_(t)R¹⁰ (where t is 0 to 2), —S(O)_(t)N(R¹⁰)₂(where t is 0 to 2), heterocyclyl and heterocyclylalkyl;

R⁴ is hydrogen, alkyl, aralkyl, aryl, haloalkyl, cycloalkyl,cycloalkylalkyl, heterocyclyl or heterocyclylalkyl;

each R⁵ is independently hydrogen, alkyl, aralkyl, or aryl;

each R⁷ is an optionally substituted alkylene chain of one carbon;

each R⁸ is an optionally substituted straight or branched alkylene oralkenylene chain of two carbons;

each R⁹ is an optionally substituted alkylene chain of three carbons;

each R¹⁰ is hydrogen, alkyl, alkenyl, cycloalkyl, cycloalkylalkyl,aralkyl or aryl; and

R¹¹ is hydrogen, alkyl or aralkyl;

as a single stereoisomer, a mixture of stereoisomers, or as a racemicmixture of stereoisomers; or as a solvate or polymorph; or as apharmaceutically acceptable salt thereof;

wherein the method comprises administering the compound of formula (I)to a mammalian cell and the compound of formula (I) is capable ofinhibiting the activity of PTPN12, PTPN2, and/or PTPN1 within themammalian cell.

In another aspect, this invention provides a pharmaceutical compositionuseful in treating cancer or inflammation in a human, wherein thepharmaceutical composition comprises a pharmaceutically acceptablecarrier, diluent or excipient and a compound of formula (Ia):

wherein:

each p is independently 1 to 5;

A is linker of four atoms and is selected from the group consisting ofan optionally substituted straight or branched alkylene chain of fourcarbons, an optionally substituted straight or branched alkenylene chainof four carbons, —R^(8a)—C(R^(2a))—N(R^(5a))—,—R^(8a)—N(R^(5a))—C(R^(2a))—, —R^(8a)—O—C(R^(2a))—,—R^(7a)-O-C(R^(2a))—R^(7a)—, —R^(8a)—O—R^(7a)—,—R^(7a)—C(R^(2a))—N(R^(5a))—S(O)_(t)— (where t is 0 to 2),—R^(8a)—N(R^(5a))—R^(7a), —R^(8a)—S(O)_(t)—R^(7a)— (where t is 0 to 2),—R^(9a)—N(R^(5a))—, —R^(9a)—O—, and —R^(9a)—C(R^(2a))—;

R^(1a) and R^(2a) are each independently ═O or ═S;

R^(3a) is alkyl, alkenyl, aryl, aralkyl, aralkenyl, cycloalkyl,cycloalkylalkyl, chloro, iodo, bromo, haloalkyl, haloalkoxy, nitro,cyano, —N═N—O—R^(11a), —OR^(10a), —C(O)OR^(10a), —C(O)N(R^(10a))₂,—N(R^(12a))₂, —N(R^(10a))C(O)R^(10a), —N(R^(10a))C(O)OR^(11a),—S(O)_(t)R^(10a) (where t is 0 to 2), —S(O)_(t)N(R^(10a))₂ (where t is 0to 2), or heterocyclylalkyl;

R^(4a) is hydrogen, alkyl, aralkyl, aryl, haloalkyl, cycloalkyl,cycloalkylalkyl, heterocyclyl or heterocyclylalkyl;

R^(5a) is hydrogen, alkyl, aralkyl, or aryl;

R^(6a) is alkyl, alkenyl, aryl, aralkyl, aralkenyl, cycloalkyl,cycloalkylalkyl, halo, haloalkyl, haloalkoxy, nitro, cyano,—N═N—O—R^(11a), —C(O)N(R^(10a))₂, —N(R^(10a))₂, —N(R^(10a))C(O)R^(10a),—N(R^(10a))C(O)OR^(11a), —S(O)_(t)R^(10a) (where t is 0 to 2),—S(O)_(t)N(R^(10a))₂ (where t is 0 to 2), heterocyclyl orheterocyclylalkyl;

each R^(7a) is an optionally substituted alkylene chain of one carbon;

each R^(8a) is an optionally substituted straight or branched alkyleneor alkenylene chain of two carbons;

each R^(9a) is an optionally substituted alkylene chain of threecarbons;

each R^(10a) is hydrogen, alkyl, alkenyl, cycloalkyl, cycloalkylalkyl,aralkyl or aryl;

R^(11a) is hydrogen, alkyl or aralkyl; and

R^(12a) is hydrogen, aryl or aralkyl;

as a single stereoisomer, a mixture of stereoisomers, or as a racemicmixture of stereoisomers; or as a solvate or polymorph; or as apharmaceutically acceptable salt thereof.

In another aspect of the invention, the use of pharmaceuticalcompositions of the invention for the treatment of cancer orinflammation in provided.

In another aspect of the invention, the use of pharmaceuticalcompositions are provided for use in treating colon or colorectalcancer.

In another aspect of the invention, the use of pharmaceuticalcompositions of the invention in the manufacture of medicaments for thetreatment of cancer and/or inflammation are provided.

In another aspect of the invention, the use of pharmaceuticalcompositions are provided for the treatment of disorders associated withhyperproliferation, tissue remodeling, and/or tissue repair.

In another aspect of the invention, the use of pharmaceuticalcompositions are provided for the treatment of endocrine disorders.

In another aspect of the invention, the use of pharmaceuticalcompositions are provided for the treatment of disorders associated withPTPN12, PTPN2, and/or PTPN1 expression.

DETAILED DESCRIPTION OF THE INVENTION

Definitions

As used herein the singular forms “a”, “and”, and “the” include pluralreferents unless the context clearly dictates otherwise. For example, “acompound” refers to one or more of such compounds, while “the enzyme”includes a particular enzyme as well as other family members andequivalents thereof as known to those skilled in the art. As used in thespecification and appended claims, unless specified to the contrary, thefollowing terms have the meaning indicated.

“Alkyl” refers to a straight or branched hydrocarbon chain radicalconsisting solely of carbon and hydrogen atoms, containing nounsaturation, having from one to eight carbon atoms, and which isattached to the rest of the molecule by a single bond, e.g., methyl,ethyl, n-propyl, 1-methylethyl(iso-propyl), n-butyl, n-pentyl,1,1-dimethylethyl(t-butyl), and the like. Unless stated otherwisespecifically in the specification, the alkyl radical may be optionallysubstituted by hydroxy, alkoxy, aryloxy, haloalkoxy, cyano, nitro,mercapto, alkylthio, —N(R⁸)₂, —C(O)OR⁸, —C(O)N(R⁸)₂ or —N(R⁸)C(O)R⁸where each R⁸ is independently hydrogen, alkyl, alkenyl, cycloalkyl,cycloalkylalkyl, aralkyl or aryl. Unless stated otherwise specificallyin the specification, it is understood that for radicals, as definedbelow, that contain a substituted alkyl group that the substitution canoccur on any carbon of the alkyl group.

“Alkoxy” refers to a radical of the formula —OR_(a) where R_(a) is analkyl radical as defined above, e.g., methoxy, ethoxy, n-propoxy,1-methylethoxy(iso-propoxy), n-butoxy, n-pentoxy,1,1-dimethylethoxy(t-butoxy), and the like. Unless stated otherwisespecifically in the specification, it is understood that for radicals,as defined below, that contain a substituted alkoxy group that thesubstitution can occur on any carbon of the alkoxy group. The alkylradical in the alkoxy radical may be optionally substituted as describedabove.

“Alkylthio” refers to a radical of the formula —SR_(a) where R_(a) is analkyl radical as defined above, e.g., methylthio, ethylthio,n-propylthio, 1-methylethylthio(iso-propylthio), n-butylthio,n-pentylthio, 1,1-dimethylethylthio(t-butylthio), and the like. Unlessstated otherwise specifically in the specification, it is understoodthat for radicals, as defined below, that contain a substitutedalkylthio group that the substitution can occur on any carbon of thealkylthio group. The alkyl radical in the alkylthio radical may beoptionally substituted as described above.

“Alkenyl” refers to a straight or branched hydrocarbon chain radicalconsisting solely of carbon and hydrogen atoms, containing at least onedouble bond, having from two to eight carbon atoms, and which isattached to the rest of the molecule by a single bond or a double bond,e.g., ethenyl, prop-1-enyl, but-1-enyl, pent-1-enyl, penta-1,4-dienyl,and the like. Unless stated otherwise specifically in the specification,the alkenyl radical may be optionally substituted by hydroxy, alkoxy,haloalkoxy, cyano, nitro, mercapto, alkylthio, cycloalkyl, —N(R⁸)₂,—C(O)OR⁸, —C(O)N(R⁸)₂ or —N(R⁸)—C(O)—R⁸ where each R⁸ is independentlyhydrogen, alkyl, alkenyl, cycloalkyl, cycloalkylalkyl, aralkyl or aryl.Unless stated otherwise specifically in the specification, it isunderstood that for radicals, as defined below, that contain asubstituted alkenyl group that the substitution can occur on any carbonof the alkenyl group.

“Aryl” refers to a phenyl or naphthyl radical. Unless stated otherwisespecifically in the specification, the term “aryl” or the prefix “ar-”(such as in “aralkyl”) is meant to include aryl radicals optionallysubstituted by one or more substituents selected from the groupconsisting of hydroxy, alkoxy, aryloxy, haloalkoxy, cyano, nitro,mercapto, alkylthio, cycloalkyl, —N(R⁸)₂, —C(O)OR⁸, —C(O)N(R⁸)₂ or—N(R⁸)C(O)R⁸ where each R⁸ is independently hydrogen, alkyl, alkenyl,cycloalkyl, cycloalkylalkyl, aralkyl or aryl.

“Aralkyl” refers to a radical of the formula —R_(a)R_(b) where R_(a) isan alkyl radical as defined above and R_(b) is one or more aryl radicalsas defined above, e.g., benzyl, diphenylmethyl and the like. The arylradical(s) may be optionally substituted as described above.

“Aralkenyl” refers to a radical of the formula —R_(c)R_(b) where R_(c)is an alkenyl radical as defined above and R_(b) is one or more arylradicals as defined above, e.g., 3-phenylprop-1-enyl, and the like. Thearyl radical(s) and the alkenyl radical may be optionally substituted asdescribed above.

“Alkylene chain” refers to a straight or branched divalent hydrocarbonchain consisting solely of carbon and hydrogen, containing nounsaturation and having from one to eight carbon atoms, e.g., methylene,ethylene, propylene, n-butylene, and the like. The alkylene chain may beoptionally substituted by one or more substituents selected from thegroup consisting of aryl, halo, hydroxy, alkoxy, haloalkoxy, cyano,nitro, mercapto, alkylthio, cycloalkyl, —N(R⁸)₂, —C(O)OR⁸, —C(O)N(R⁸)₂or —N(R⁸)C(O)R⁸ where each R⁸ is independently hydrogen, alkyl, alkenyl,cycloalkyl, cycloalkylalkyl, aralkyl or aryl. The alkylene chain may beattached to the rest of the molecule through any two carbons within thechain.

“Alkenylene chain” refers to a straight or branched divalent hydrocarbonchain consisting solely of carbon and hydrogen, containing at least onedouble bond and having from two to eight carbon atoms, e.g., ethenylene,prop-1-enylene, but-1-enylene, pent-1-enylene, hexa-1,4-dienylene, andthe like. The alkenylene chain may be optionally substituted by one ormore substituents selected from the group consisting of aryl, halo,hydroxy, alkoxy, haloalkoxy, cyano, nitro, mercapto, alkylthio,cycloalkyl, —N(R⁸)₂, —C(O)OR⁸, —C(O)N(R⁸)₂ or —N(R⁸)C(O)R⁸ where each R⁸is independently hydrogen, alkyl, alkenyl, cycloalkyl, cycloalkylalkyl,aralkyl or aryl. The alkenylene chain may be attached to the rest of themolecule through any two carbons within the chain.

“Cycloalkyl” refers to a stable monovalent monocyclic or bicyclichydrocarbon radical consisting solely of carbon and hydrogen atoms,having from three to ten carbon atoms, and which is saturated andattached to the rest of the molecule by a single bond, e.g.,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, decalinyl and thelike. Unless otherwise stated specifically in the specification, theterm “cycloalkyl” is meant to include cycloalkyl radicals which areoptionally substituted by one or more substituents independentlyselected from the group consisting of alkyl, aryl, aralkyl, halo,haloalkyl, hydroxy, alkoxy, haloalkoxy, cyano, nitro, mercapto,alkylthio, cycloalkyl, —N(R⁸)₂, —C(O)OR⁸, —C(O)N(R⁸)₂ or —N(R⁸)C(O)R⁸where each R⁸ is independently hydrogen, alkyl, alkenyl, cycloalkyl,cycloalkylalkyl, aralkyl or aryl.

“Cycloalkylalkyl” refers to a radical of the formula —R_(a)R_(d) whereR_(a) is an alkyl radical as defined above and R_(d) is a cycloalkylradical as defined above. The alkyl radical and the cycloalkyl radicalmay be optionally substituted as defined above.

“Halo” refers to bromo, chloro, fluoro or iodo.

“Haloalkyl” refers to an alkyl radical, as defined above, that issubstituted by one or more halo radicals, as defined above, e.g.,trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl,1-fluoromethyl-2-fluoroethyl, 3-bromo-2-fluoropropyl,1-bromomethyl-2-bromoethyl, and the like.

“Haloalkoxy” refers to a radical of the formula —OR_(c) where R_(c) isan haloalkyl radical as defined above, e.g., trifluoromethoxy,difluoromethoxy, trichloromethoxy, 2,2,2-trifluoroethoxy,1-fluoromethyl-2-fluoroethoxy, 3-bromo-2-fluoropropoxy,1-bromomethyl-2-bromoethoxy, and the like.

“Heterocyclyl” refers to a stable 3- to 15-membered ring radical whichconsists of carbon atoms and from one to five heteroatoms selected fromthe group consisting of nitrogen, oxygen and sulfur. For purposes ofthis invention, the heterocyclyl radical may be a monocyclic, bicyclicor tricyclic ring system, which may include fused or bridged ringsystems; and the nitrogen, carbon or sulfur atoms in the heterocyclylradical may be optionally oxidized; the nitrogen atom may be optionallyquaternized; and the heterocyclyl radical may be aromatic or partiallyor fully saturated. The heterocyclyl radical may not be attached to therest of the molecule at any heteroatom atom. Examples of suchheterocyclyl radicals include, but are not limited to, azepinyl,acridinyl, benzimidazolyl, benzthiazolyl, benzothiadiazolyl,benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl,benzopyranonyl, benzofuranyl, benzofuranonyl,benzothienyl(benzothiophenyl), benzotriazolyl, carbazolyl, cinnolinyl,decahydroisoquinolyl, dioxolanyl, furanyl, furanonyl, isothiazolyl,imidazolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, indolyl,indazolyl, isoindolyl, indolinyl, isoindolinyl, indolizinyl, isoxazolyl,isoxazolidinyl, morpholinyl, naphthyridinyl, oxadiazolyl,octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl,2-oxopiperidinyl, 2-oxopyrrolidinyl, 2-oxoazepinyl, oxazolyl,oxazolidinyl, oxiranyl, piperidinyl, piperazinyl, 4-piperidonyl,phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl,purinyl, pyrrolyl, pyrrolidinyl, pyrazolyl, pyrazolidinyl, pyridinyl,pyrazinyl, pyrimidinyl, pyridazinyl, quinazolinyl, quinoxalinyl,quinolinyl, quinuclidinyl, isoquinolinyl, thiazolyl, thiazolidinyl,thiadiazolyl, triazolyl, tetrazolyl, tetrahydrofuryl, triazinyl,tetrahydropyranyl, thienyl, thiamorpholinyl, thiamorpholinyl sulfoxide,and thiamorpholinyl sulfone. Unless stated otherwise specifically in thespecification, the term “heterocyclyl” is meant to include heterocyclylradicals as defined above which are optionally substituted by one ormore substituents selected from the group consisting of alkyl, halo,nitro, cyano, haloalkyl, haloalkoxy, aryl, heterocyclyl,heterocyclylalkyl, —OR⁸, —R⁷—OR⁸, —C(O)OR⁸, —R⁷—C(O)OR⁸, —C(O)N(R⁸)₂,—N(R⁸)₂, —R⁷—N(R⁸)₂, and —N(R⁸)C(O)R⁸ wherein each R⁷ is a straight orbranched alkylene or alkenylene chain and each R⁸ is independentlyhydrogen, alkyl, alkenyl, cycloalkyl, cycloalkylalkyl, aralkyl or aryl.

“Heterocyclylalkyl” refers to a radical of the formula —R_(a)R_(e) whereR_(a) is an alkyl radical as defined above and R_(e), is a heterocyclylradical as defined above, and if the heterocyclyl is anitrogen-containing heterocyclyl, the heterocyclyl may be attached tothe alkyl radical at the nitrogen atom. The heterocyclyl radical may beoptionally substituted as defined above.

As used herein, compounds which are “commercially available” may beobtained from standard commercial sources including Acros Organics(Pittsburgh, Pa.), Aldrich Chemical (Milwaukee Wis., including SigmaChemical and Fluka), Apin Chemicals Ltd. (Milton Park, UK), AvocadoResearch (Lancashire, U.K.), BDH Inc. (Toronto, Canada), Bionet(Cornwall, U.K.), Chemservice Inc. (West Chester, Pa.), CrescentChemical Co. (Hauppauge, N.Y.), Eastman Organic Chemicals, Eastman KodakCompany (Rochester, N.Y.), Fisher Scientific Co. (Pittsburgh, Pa.),Fisons Chemicals (Leicestershire, UK), Frontier Scientific (Logan,Utah), ICN Biomedicals, Inc. (Costa Mesa, Calif.), Key Organics(Cornwall, U.K), Lancaster Synthesis (Windham, N.H.), Maybridge ChemicalCo. Ltd. (Cornwall, U.K.), Parish Chemical Co. (Orem, Utah), Pfaltz &Bauer, Inc. (Waterbury, Conn.), Polyorganix (Houston, Tex.), PierceChemical Co. (Rockford, Ill.), Riedel de Haen AG (Hannover, Germany),Spectrum Quality Product, Inc. (New Brunswick, N.J.), TCI America(Portland, Oreg.), Trans World Chemicals, Inc. (Rockville, Md.), andWako Chemicals USA, Inc. (Richmond, Va.).

As used herein, “suitable conditions” for carrying out a synthetic stepare explicitly provided herein or may be discerned by reference topublications directed to methods used in synthetic organic chemistry.The reference books and treatise set forth above that detail thesynthesis of reactants useful in the preparation of compounds of thepresent invention, will also provide suitable conditions for carryingout a synthetic step according to the present invention.

As used herein, “methods known to one of ordinary skill in the art” maybe identified though various reference books and databases. Suitablereference books and treatise that detail the synthesis of reactantsuseful in the preparation of compounds of the present invention, orprovide references to articles that describe the preparation, includefor example, “Synthetic Organic Chemistry”, John Wiley & Sons, Inc., NewYork; S. R. Sandier et al., “Organic Functional Group Preparations,” 2ndEd., Academic Press, New York, 1983; H. O. House, “Modern SyntheticReactions”, 2nd Ed., W. A. Benjamin, Inc. Menlo Park, Calif. 1972; T. L.Gilchrist, “Heterocyclic Chemistry”, 2nd Ed., John Wiley & Sons, NewYork, 1992; J. March, “Advanced Organic Chemistry: Reactions, Mechanismsand Structure”, 4th Ed., Wiley-Interscience, New York, 1992. Specificand analogous reactants may also be identified through the indices ofknown chemicals prepared by the Chemical Abstract Service of theAmerican Chemical Society, which are available in most public anduniversity libraries, as well as through on-line databases (the AmericanChemical Society, Washington, D.C., www.acs.org may be contacted formore details). Chemicals that are known but not commercially availablein catalogs may be prepared by custom chemical synthesis houses, wheremany of the standard chemical supply houses (e.g., those listed above)provide custom synthesis services.

“Prodrugs” is meant to indicate a compound that may be converted underphysiological conditions or by solvolysis to a biologically activecompound of the invention. Thus, the term “prodrug” refers to ametabolic precursor of a compound of the invention that ispharmaceutically acceptable. A prodrug may be inactive when administeredto a subject in need thereof, but is converted in vivo to an activecompound of the invention. Prodrugs are typically rapidly transformed invivo to yield the parent compound of the invention, for example, byhydrolysis in blood. The prodrug compound often offers advantages ofsolubility, tissue compatibility or delayed release in a mammalianorganism (see, Bundgard, H., Design of Prodrugs (1985), pp. 7-9, 21-24(Elsevier, Amsterdam).

A discussion of prodrugs is provided in Higuchi, T., et al., “Pro-drugsas Novel Delivery Systems,” A.C.S. Symposium Series, Vol. 14, and inBioreversible Carriers in Drug Design, ed. Edward B. Roche, AmericanPharmaceutical Association and Pergamon Press, 1987, both of which areincorporated in full by reference herein.

The term “prodrug” is also meant to include any covalently bondedcarriers which release the active compound of the invention in vivo whensuch prodrug is administered to a mammalian subject. Prodrugs of acompound of the invention may be prepared by modifying functional groupspresent in the compound of the invention in such a way that themodifications are cleaved, either in routine manipulation or in vivo, tothe parent compound of the invention. Prodrugs include compounds of theinvention wherein a hydroxy, amino or mercapto group is bonded to anygroup that, when the prodrug of the compound of the invention isadministered to a mammalian subject, cleaves to form a free hydroxy,free amino or free mercapto group, respectively. Examples of prodrugsinclude, but are not limited to, acetate, formate and benzoatederivatives of alcohol and amine functional groups in the compounds ofthe invention and the like.

“Stable compound” and “stable structure” are meant to indicate acompound that is sufficiently robust to survive isolation to a usefuldegree of purity from a reaction mixture, and formulation into anefficacious therapeutic agent.

“Mammal” includes humans and domestic animals, such as cats, dogs,swine, cattle, sheep, goats, horses, rabbits, and the like.

“Optional” or “optionally means that the subsequently described event ofcircumstances may or may not occur, and that the description includesinstances where said event or circumstance occurs and instances in whichit does not. For example, “optionally substituted aryl” means that thearyl radical may or may not be substituted and that the descriptionincludes both substituted aryl radicals and aryl radicals having nosubstitution.

“Pharmaceutically acceptable carrier, diluent or excipient” includeswithout limitation any adjuvant, carrier, excipient, glidant, sweeteningagent, diluent, preservative, dye/colorant, flavor enhancer, surfactant,wetting agent, dispersing agent, suspending agent, stabilizer, isotonicagent, solvent, or emulsifier which has been approved by the UnitedStates Food and Drug Administration as being acceptable for use inhumans or domestic animals.

“Pharmaceutically acceptable salt” includes both acid and base additionsalts.

“Pharmaceutically acceptable acid addition salt” refers to those saltswhich retain the biological effectiveness and properties of the freebases, which are not biologically or otherwise undesirable, and whichare formed with inorganic acids such as hydrochloric acid, hydrobromicacid, sulfuric acid, nitric acid, phosphoric acid and the like, andorganic acids such as acetic acid, trifluoroacetic acid, propionic acid,glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid,succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid,cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid,p-toluenesulfonic acid, salicylic acid, and the like.

“Pharmaceutically acceptable base addition salt” refers to those saltswhich retain the biological effectiveness and properties of the freeacids, which are not biologically or otherwise undesirable. These saltsare prepared from addition of an inorganic base or an organic base tothe free acid. Salts derived from inorganic bases include, but are notlimited to, the sodium, potassium, lithium, ammonium, calcium,magnesium, iron, zinc, copper, manganese, aluminum salts and the like.Preferred inorganic salts are the ammonium, sodium, potassium, calcium,and magnesium salts. Salts derived from organic bases include, but arenot limited to, salts of primary, secondary, and tertiary amines,substituted amines including naturally occurring substituted amines,cyclic amines and basic ion exchange resins, such as isopropylamine,trimethylamine, diethylamine, triethylamine, tripropylamine,ethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol,dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine,hydrabamine, choline, betaine, ethylenediamine, glucosamine,methylglucamine, theobromine, purines, piperazine, piperidine,N-ethylpiperidine, polyamine resins and the like. Particularly preferredorganic bases are isopropylamine, diethylamine, ethanolamine,trimethylamine, dicyclohexylamine, choline and caffeine.

“PTPN12” refers to the Human Genome Organization (HUGO) NomenclatureCommittee's name for protein tyrosine phosphatase, non-receptor like 12.PTPN12 is also known as PTP-PEST and PTPG1.

“PTPN1” refers to the HUGO Nomenclature Committee's name for proteintyrosine phosphatase, non-receptor like 1. PTPN1 is also known as PTP1B.

“PTPN2” refers to the Human Genome Organization (HUGO) NomenclatureCommitee's name for protein tyrosine phosphatase, non-receptor like 2.PTPN2 is also known as TC-PTP or T-cell-PTP. The sequence of PTPN2 maybe accessed at Genbank, M25393, and is described in Cool et al. (1989)Proc. Natl. Acad. Sci. U.S.A. 86 (14), 5257-5261.

“Therapeutically effective amount” refers to that amount of a compoundof formula (I) which, when administered to a mammal, preferably a human,is sufficient to effect treatment, as defined below, for cancer andinflammation in the mammal. The amount of a compound of formula (I)which constitutes a “therapeutcally effective amount” will varydepending on the compound, the condition and its severity, and the ageof the mammal to be treated, but can be determined routinely by one ofordinary skill in the art having regard to his own knowledge and to thisdisclosure.

“Treating” or “treatment” as used herein covers the treatment of cancer,inflammation, or diabetes, preferably cancer or inflammation associatedwith PTPN12 or PTPN2 activity, or diabetes associated with PTPN1activity, in a mammal, preferably a human, and includes:

(i) preventing the cancer, inflammation, or diabetes from occurring in amammal, in particular, when such mammal is predisposed to the conditionbut has not yet been diagnosed as having it;

(ii) inhibiting the cancer, inflammation or diabetes i.e., arresting itsdevelopment; or

(iii) relieving the cancer, inflammation or diabetes, i.e., causingregression of the condition.

“Insulin resistance” includes diabetes, hyperglycemia, and otherdisorders associated with insulin receptor (IR) signal transduction.

The compounds of formula (I), or their pharmaceutically acceptable saltsmay contain one or more asymmetric centers and may thus give rise toenantiomers, diastereomers, and other stereoisomeric forms that may bedefined, in terms of absolute stereochemistry, as (R)- or (S)- or, as(D)- or (L)- for amino acids. The present invention is meant to includeall such possible isomers, as well as, their racemic and optically pureforms. Optically active (+) and (−), (R)- and (S)—, or (D)- and(L)-isomers may be prepared using chiral synthons or chiral reagents, orresolved using conventional techniques, such as reverse phase HPLC. Whenthe compounds described herein contain olefinic double bonds or othercenters of geometric asymmetry, and unless specified otherwise, it isintended that the compounds include both E and Z geometric isomers.Likewise, all tautomeric forms are also intended to be included.

The nomenclature used herein for the compounds of formula (I) is amodified form of the I.U.P.A.C. nomenclature system wherein thecompounds are named herein as derivatives of the thiazolidine moiety.

Methods of Use

This invention is directed to methods of using compounds of formula (I)and formula (Ia), as set forth above in the Summary of the Invention,and pharmaceutical compositions containing compounds of formula (Ia) intreating diseases of cell hyperproliferation and activation, includingcancer and, inflammation.

The methods of the invention can be used prophylactically (i.e., toprevent the disorder of interest from occurring) or therapeutically(i.e., to inhibit or relieve the disorder). As used herein, the term“treating” is used to refer to both prevention of disease, and treatmentof pre-existing conditions. The prevention of symptoms is accomplishedby administration of the compounds and pharmaceutical compositions ofthe invention prior to development of overt disease, e.g., to preventthe regrowth of tumors, prevent metastatic growth, diminish restenosisassociated with cardiovascular surgery, to prevent or reduce cellmigration leading to inflammation and associated tissue damage.Alternatively, the compounds and pharmaceutical compositions of theinvention may be administered to a subject in need thereof to treat anongoing disease, by stabilizing or improving the clinical symptoms ofthe patient.

The subject, or patient, may be from any mammalian species, e.g.primates, particularly humans; rodents, including mice, rats andhamsters; rabbits; equines; bovines; canines; felines; etc. Animalmodels are of interest for experimental investigations, providing amodel for treatment of human disease.

The susceptibility of a particular cell to treatment according to theinvention may be determined by in vitro testing. Typically, a culture ofthe cell is combined with a subject compound at varying concentrationsfor a period of time sufficient to allow the active agents to inducecell death or inhibit migration, usually between about one hour and oneweek. For in vitro testing, cultured cells from a biopsy sample may beused.

The dose will vary depending on mode of administration, specificdisorder, patient status, etc. Typically a therapeutic dose will besufficient to substantially decrease the undesirable cell population inthe targeted tissue, while maintaining patient viability. Treatment willgenerally be continued until there is a substantial reduction, e.g. atleast about 50%, decrease in the clinical manifestation of disease, andmay be continued until there are essentially none of the undesirablecellular activity detected in the relevant tissue.

The compounds of formula (I) and the compounds of formula (Ia) may alsofind use in the specific inhibition of signaling pathways mediated byprotein tyrosine phosphatases, for example, PTPN12, PTPN2, or PTPNL andas a “positive” control in high throughput screening for othermodulating compounds.

The compounds of formula (I) and the compounds of formula (Ia) may alsofind use as affinity reagents for the isolation and/or purification ofphosphatases using the biochemical affinity of the enzyme for inhibitorsthat act on it. The compounds are coupled to a matrix or gel. Thecoupled support is then used to separate the enzyme, which binds to thecompound, from a sample mixture, e.g., a cell lysate, which may beoptionally partially purified. The sample mixture is contacted with thecompound coupled support under conditions that minimize non-specificbinding. Methods known in the art include columns, gels, capillaries,etc. The unbound proteins are washed free of the resin and the boundproteins are then eluted in a suitable buffer.

The compounds of formula (I) and the compounds of formula (Ia) may alsobe useful as reagents for studying signal transduction or any of theclinical disorders listed throughout this application, and for use as apositive control in high throughput screening.

Disorders and Conditions of Interest

The conditions and disorders of interest to the present invention arecancer and inflammation, in particular, cancer and inflammationassociated with PTPN12 and/or PTPN2 activity. There are many disordersassociated with PTPN12 and/or PTPN2 activity including dysregulation ofcellular division. Accordingly, the compounds and pharmaceuticalcompositions of the invention may be used to treat a variety ofconditions where there is proliferation and/or migration of smoothmuscle cells, and/or inflammatory cells into the intimal layer of avessel, resulting in restricted blood flow through that vessel, i.e.neointimal occlusive lesions. Occlusive vascular conditions of interestinclude atherosclerosis, graft coronary vascular disease aftertransplantation, vein graft stenosis, peri-anastomatic prosthetic graftstenosis, restenosis after angioplasty or stent placement, and the like.

Disorders and conditions where there is hyperproliferation and/or tissueremodelling or repair of reproductive tissue, e.g. uterine, testicularand ovarian carcinomas, endometriosis, squamous and glandular epithelialcarcinomas of the cervix, etc. are reduced in cell number byadministration of the compounds and pharmaceutical compositions of theinvention.

Tumors of interest for treatment include carcinomas, e.g. colon,duodenal, prostate, breast, melanoma, ductal, hepatic, pancreatic,renal, endometrial, stomach, invasive oral cancer, transitional andsquamous cell urinary carcinoma etc; hematological malignancies, e.g.childhood acute leukaemia, non-Hodgkin's lymphomas, or chroniclymphocytic leukaemia, non-small cell lung carcinoma, adenocarcinoma,and melanoma. Some cancers of particular interest include breastcancers, wherein ductal carcinoma in situ is the most common type ofnoninvasive breast cancer.

Other disorders and conditions of interest relate to epidermalhyperproliferation, tissue remodelling and repair. For example, thechronic skin inflammation of psoriasis is associated with hyperplasticepidermal keratinocytes.

In one aspect of the invention, compounds and pharmaceuticalcompositions of the invention may be used to inhibit the activity ofPTPN12 and/or PTPN2 for the treatment of inflammatory disorders andautoimmune conditions including, but not limited to, psoriasis,rheumatoid arthritis, multiple sclerosis, scleroderma, systemic lupuserythematosus, Sjögren's syndrome, atopic dermatitis, asthma, andallergy. Target cells susceptible to the treatment include cellsinvolved in instigating autoimmune reactions as well as those sufferingor responding from the effects of autoimmune attack or inflammatoryevents, and include lymphocytes and fibroblasts.

PTPN12 contains a proline rich motif at its C-terminal and can bind top130^(cas), which is a focal adhesion associated protein containing anSH3 domain. In normal cells, p130^(cas) becomes highly phosphorylatedfollowing integrin dependent activation of the fak and src kinases. Thisphosphorylation appears to allow tyrosine dependent signalling that hasas a consequence the reorganization of actin filaments. Because of theimportance of integrin signalling in the cell cytoskeleton, motility andtransformation, the action of PTPN12 on p130^(cas) may have dramaticconsequences in mammalian development as well as in somephysiopathological events. The process of cell migration is crucial forthe correct development of a mammalian embryo. In an adult organism,cell migration plays an important role in events like invasion of awounded space by fibroblasts and endothelial cells and translocation oflymphocytes and neutrophiles to an inflammation site. In cancer, tumorcells also have to migrate in order to reach the circulatory system anddisperse throughout the organism. Takekawa, M. et al., FEBS Lett.(1994),Vol. 339, pp. 222-228 discloses aberrant transcripts of PTPN12 in cancercells. The effect of PTPN12 levels on fibroblast motility is describedin Garton et al. (1999) J. Biol. Chem. 274(6):3811-3818. Davidson et al.(2001) EMBO. J. 20(13):3414-26 discusses a connection of PTPN12 withinflammation. The relationship between PTPN12 and podocyte regulation inkidney is described in Reiser, J. et al., Rapid Communication, KidneyInt. (2000), Vol. 57, No. 5, pp. 2035-2042.

PTPN12 is involved in signalling pathways for such important cellularactivities as responses to extracellular signals and cell cyclecheckpoints. Inhibition of PTPN12 provides a means (for example, byblocking the effect of an extracellular signal) of intervening in thesesignalling pathways, which are associated with a variety of pathologicalor clinical conditions. PTPN12 is associated with cell adhesion, celldivision and cell migration and thus is implicated in cancer andinflammation.

Another PTP of particular interest is PTPN2. PTPN2 is also known asT-cell protein tyrosine phosphatase (TC-PTP) and was first identified byCool et al., Proc. Natl. Acad. Sci. (1989), Vol. 86, pp. 5257-5261.PTPN2 exists in two forms generated by alternative splicing: a 48 kDaendoplasmic reticular (ER)-associated form called TC48 (PTP-S4); and a45-kDa nuclear form called TC45 (PTP-S2). PTPN2 plays a significant rolein both hematopoiesis and immune function. You-Ten et al., J. Exp. Med.(1997), Vol. 186, No. 5, pp. 683-693 found that PTPN2−/− mice diebetween 3-5 weeks of age, exhibiting specific defects in bone marrow(BM), B cell lymphopoeisis, and erythropoiesis, as well as impaired Tand B cell functions. Bone marrow transplantation experimentsdemonstrated that hematopoletic failure in the homozygotes was not dueto a stem cell defect but rather stromal cell deficiency.

PTPN2 may play a role in cancer progression and metastases. Mitra, S. K.et al., Exp. Cell Res. 15 (2001), Vol. 270, No. 1, pp. 32-44demonstrated inhibition of anchorage-independent cell growth, adhesion,and cyclin D1 gene expression by a dominant negative mutant PTPN2.Expression of mutant PTPN2 in PyF cells resulted in strong inhibition ofanchorage-independent growth in soft agar but had no significant effecton growth in liquid culture. Tumor formation in nude mice was alsoreduced by the presence of mutant PTPN2.

PTPN2 plays a role in apoptosis, making it a useful target for cancertherapy or as a component of a cancer therapeutic cocktail. Zsigmond, E.et al., FEBS Lett. (1999), Vol. 453, No. 3, pp. 308-312, found thatoverexpression of PTPN2 induced nuclear fragmentation typical ofapoptosis. In addition, PTPN2 appears to be active in progressing theearly G1 phase of the cell cycle through the NF-kappaB pathway(Ibarra-Sanches, M. J. et al., Oncogene (2001), Vol. 20, No. 34, pp.4728-39). Inhibition of PTPN2 is useful in treating conditionsassociated with PTPN2 activity, such as inflammation, cancer progressionand metastases.

PTPN1 activity is associated with insulin resistance, and diabetes,hyperglycemia, and other disorders associated with insulin receptor (IR)signal transduction. Reduction of PTPN1, for example, is sufficient toincrease insulin-dependent metabolic signaling and improve insulinsensitivity (Gum, R. J.; Gaede, L. L.; Koterski, S. L. et al., Diabetes(2003) 52(1):21-8). When PTPN1 is overexpressed, it plays a role ininsulin resistance (Ahmad, F. et al., (1997) J. Clin. Invest. 100:449-458, 1997).

Administration of the Compounds and Pharmaceutical Compositions of theInvention

Administration of the compounds of the invention, or theirpharmaceutically acceptable salts, in pure form or in an appropriatepharmaceutical composition, can be carried out via any of the acceptedmodes of administration of agents for serving similar utilities. Thepharmaceutical compositions of the invention can be prepared bycombining a compound of the invention with an appropriatepharmaceutically acceptable carrier, diluent or excipient, and may beformulated into preparations in solid, semi-solid, liquid or gaseousforms, such as tablets, capsules, powders, granules, ointments,solutions, suppositories, injections, inhalants, gels, microspheres, andaerosols. Typical routes of administering such pharmaceuticalcompositions include, without limitation, oral, topical, transdermal,inhalation, parenteral, sublingual, rectal, vaginal, and intranasal. Theterm parenteral as used herein includes subcutaneous injections,intravenous, intramuscular, intrasternal injection or infusiontechniques. Pharmaceutical compositions of the invention are formulatedso as to allow the active ingredients contained therein to bebioavailable upon administration of the composition to a patient.Compositions that will be administered to a subject or patient take theform of one or more dosage units, where for example, a tablet may be asingle dosage unit, and a container of a compound of the invention inaerosol form may hold a plurality of dosage units. Actual methods ofpreparing such dosage forms are known, or will be apparent, to thoseskilled in this art; for example, see Remington's PharmaceuticalSciences, 18th Ed., (Mack Publishing Company, Easton, Pa., 1990). Thecomposition to be administered will, in any event, contain atherapeutically effective amount of a compound of the invention, or apharmaceutically acceptable salt thereof, for treatment of a disorder orcondition associated with hyperproliferation and tissue remodelling orrepair in accordance with the teachings of this invention.

A pharmaceutical composition of the invention may be in the form of asolid or liquid. In one aspect, the carrier(s) are particulate, so thatthe compositions are, for example, in tablet or powder form. Thecarrier(s) may be liquid, with the compositions being, for example, anoral syrup, injectable liquid or an aerosol, which is useful in, e.g.,inhalatory administration.

When intended for oral administration, the pharmaceutical composition ispreferably in either solid or liquid form, where semi-solid,semi-liquid, suspension and gel forms are included within the formsconsidered herein as either solid or liquid.

As a solid composition for oral administration, the pharmaceuticalcomposition may be formulated into a powder, granule, compressed tablet,pill, capsule, chewing gum, wafer or the like form. Such a solidcomposition will typically contain one or more inert diluents or ediblecarriers. In addition, one or more of the following may be present:binders such as carboxymethylcellulose, ethyl cellulose,microcrystalline cellulose, gum tragacanth or gelatin; excipients suchas starch, lactose or dextrins, disintegrating agents such as alginicacid, sodium alginate, Primogelm, corn starch and the like; lubricantssuch as magnesium stearate or Sterotex™; glidants such as colloidalsilicon dioxide; sweetening agents such as sucrose or saccharin; aflavoring agent such as peppermint, methyl salicylate or orangeflavoring; and a coloring agent.

When the pharmaceutical composition is in the form of a capsule, e.g., agelatin capsule, it may contain, in addition to materials of the abovetype, a liquid carrier such as polyethylene glycol or a fatty oil.

The pharmaceutical composition may be in the form of a liquid, e.g., anelixir, syrup, solution, emulsion or suspension. The liquid may be fororal administration or for delivery by injection, as two examples. Whenintended for oral administration, preferred composition contain, inaddition to the present compounds, one or more of a sweetening agent,preservatives, dye/colorant and flavor enhancer. In a compositionintended to be administered by injection, one or more of a surfactant,preservative, wetting agent, dispersing agent, suspending agent, buffer,stabilizer and isotonic agent may be included.

The liquid pharmaceutical compositions of the invention, whether they besolutions, suspensions or other like form, may include one or more ofthe following adjuvants: sterile diluents such as water for injection,saline solution, preferably physiological saline, Ringer's solution,isotonic sodium chloride, fixed oils such as synthetic mono ordiglycerides which may serve as the solvent or suspending medium,polyethylene glycols, glycerin, propylene glycol or other solvents;antibacterial agents such as benzyl alcohol or methyl paraben;antioxidants such as ascorbic acid or sodium bisulfite; chelating agentssuch as ethylenediaminetetraacetic acid; buffers such as acetates,citrates or phosphates and agents for the adjustment of tonicity such assodium chloride or dextrose. The parenteral preparation can be enclosedin ampoules, disposable syringes or multiple dose vials made of glass orplastic. Physiological saline is a preferred adjuvant. An injectablepharmaceutical composition is preferably sterile.

A liquid pharmaceutical composition of the invention intended for eitherparenteral or oral administration should contain an amount of a compoundof the invention such that a suitable dosage will be obtained.Typically, this amount is at least 0.01% of a compound of the inventionin the composition. When intended for oral administration, this amountmay be varied to be between 0.1 and about 70% of the weight of thecomposition. Preferred oral pharmaceutical compositions contain betweenabout 4% and about 50% of the compound of the invention. Preferredpharmaceutical compositions and preparations according to the presentinvention are prepared so that a parenteral dosage unit contains between0.01 to 1% by weight of the compound of the invention.

The pharmaceutical composition of the invention may be intended fortopical administration, in which case the carrier may suitably comprisea solution, emulsion, ointment or gel base. The base, for example, maycomprise one or more of the following: petrolatum, lanolin, polyethyleneglycols, bee wax, mineral oil, diluents such as water and alcohol, andemulsifiers and stabilizers. Thickening agents may be present in apharmaceutical composition for topical administration. If intended fortransdermal administration, the composition may include a transdermalpatch or iontophoresis device. Topical formulations may contain aconcentration of the compound of the invention from about 0.1 to about10% w/v (weight per unit volume).

The pharmaceutical composition of the invention may be intended forrectal administration, in the form, e.g., of a suppository, which willmelt in the rectum and release the drug. The composition for rectaladministration may contain an oleaginous base as a suitablenonirritating excipient. Such bases include, without limitation,lanolin, cocoa butter and polyethylene glycol.

The pharmaceutical composition of the invention may include variousmaterials, which modify the physical form of a solid or liquid dosageunit. For example, the composition may include materials that form acoating shell around the active ingredients. The materials that form thecoating shell are typically inert, and may be selected from, forexample, sugar, shellac, and other enteric coating agents.Alternatively, the active ingredients may be encased in a gelatincapsule.

The pharmaceutical composition of the invention in solid or liquid formmay include an agent that binds to the compound of the invention andthereby assists in the delivery of the compound. Suitable agents thatmay act in this capacity include a monoclonal or polyclonal antibody, aprotein or a liposome.

The pharmaceutical composition of the invention may consist of dosageunits which can be administered as an aerosol. The term aerosol is usedto denote a variety of systems ranging from those of colloidal nature tosystems consisting of pressurized packages. Delivery may be by aliquefied or compressed gas or by a suitable pump system that dispensesthe active ingredients. Aerosols of compounds of the invention may bedelivered in single phase, bi-phasic, or tri-phasic systems in order todeliver the active ingredient(s). Delivery of the aerosol includes thenecessary container, activators, valves, subcontainers, and the like,which together may form a kit. One skilled in the art, without undueexperimentation may determine preferred aerosols.

Whether in solid, liquid or gaseous form, the pharmaceutical compositionof the present invention may contain one or more known pharmacologicalagents used in the treatment of cancer or inflammation in a mammal,particularly, cancer or inflammation associated with hyperproliferationand tissue remodelling or repair.

The pharmaceutical compositions of the invention may be prepared bymethodology well known in the pharmaceutical art. For example, apharmaceutical composition intended to be administered by injection canbe prepared by combining a compound of the invention with water so as toform a solution. A surfactant may be added to facilitate the formationof a homogeneous solution or suspension. Surfactants are compounds thatnon-covalently interact with the compound of the invention so as tofacilitate dissolution or homogeneous suspension of the compound in theaqueous delivery system.

The compounds of the invention, or their pharmaceutically acceptablesalts, are administered in a therapeutically effective amount, whichwill vary depending upon a variety of factors including the activity ofthe specific compound employed; the metabolic stability and length ofaction of the compound; the age, body weight, general health, sex, anddiet of the patient; the mode and time of administration; the rate ofexcretion; the drug combination; the severity of the particular disorderor condition; and the subject undergoing therapy. Generally, atherapeutically effective daily dose is from about 0.1 mg to about 20mg/kg of body weight per day of a compound of the invention, or apharmaceutically acceptable salt thereof; preferably, from about 0.1 mgto about 10 mg/kg of body weight per day; and most preferably, fromabout 0.1 mg to about 7.5 mg/kg of body weight per day.

PREFERRED EMBODIMENTS OF THE INVENTION

Of the methods of using compounds of formula (I) or formula (1a) totreat cancer, inflammation, or insulin resistance in a mammal as setforth above in the Summary of the Invention, a preferred group ofmethods is that group wherein the mammal is human. Of this preferredgroup, a preferred subgroup of methods is that subgroup wherein thecancer is colorectal, or associated with hyperproliferation or tissueremodelling or repair. Of this preferred subgroup, a preferred class ofmethods is that class wherein the cancer is associated the activity ofPTPN12, PTPN1 and/or PTPN2.

Of this preferred group, subgroup and class of methods set forth above,a preferred subclass of methods is that subclass wherein the compound offormula (I) is a compound of formula (I) wherein:

A is —R⁸—C(R²)—N(R⁵)—;

R¹ and R² are each independently ═O or ═S;

R³ and R⁶ are each independently selected from the group consisting ofhydrogen, alkyl, aryl, aralkyl, halo, haloalkyl, haloalkoxy, nitro,cyano, —N═N—O—R¹¹—, —OR¹⁰, —C(O)OR¹⁰, —C(O)N(R¹⁰)₂, —N(R¹⁰)₂,—N(R¹⁰)C(O)R¹⁰, —N(R¹⁰)C(O)OR¹¹, —S(O)_(t)R¹⁰ (where t is 0 to 2),—S(O)_(t)N(R¹⁰)₂ (where t is 0 to 2), and —S(O)_(t)NH—R¹⁴;

R⁴ is hydrogen or alkyl;

R⁵ is hydrogen or alkyl;

R⁸ is an optionally substituted straight or branched alkylene oralkenylene chain of two carbons;

each R¹⁰ is hydrogen, alkyl, aralkyl or aryl; and

R¹¹ is hydrogen, alkyl or aralkyl; and

R¹⁴ is a thiazole.

Of this preferred subclass of methods, a preferred set of methods isthat set wherein the compound of formula (I) is a compound of formula(I) wherein:

A is —R⁸—C(R²)—N(R⁵)—;

R¹ and R² are each independently ═O or ═S;

R³ is hydrogen, alkyl, alkoxy, aryl, aralkyl, halo, haloalkyl, orhaloalkoxy;

R⁴ is hydrogen or alkyl;

R⁵ is hydrogen or alkyl;

R⁶ is —C(O)OR¹⁰, —C(O)N(R¹⁰)₂, —S(O)_(t)R¹⁰ (where t is 0 to 2),—S(O)_(t)N(R¹⁰)₂ (where t is 0 to 2), oe and —S(O)_(t)NH—R¹⁴;

R⁸ is an optionally substituted straight or, branched alkylene oralkenylene chain of two carbons; and

each R¹⁰ is hydrogen, alkyl, aralkyl or aryl.

Of this set of methods, a preferred subset of methods is that subsetwherein the compound of formula (I) is a compound of formula (I)wherein:

A is —R⁸—C(R²)—N(R⁵)—;

R¹ and R² are both ═O;

R³ is alkyl, alkoxy, haloalkyl or haloalkoxy;

R⁴ is hydrogen;

R⁵ is hydrogen or alkyl;

R⁶ is —S(O)₂N(R¹⁰)₂;

R⁸ is ethylene;

R¹⁰ is hydrogen or alkyl; and

R¹⁴ is a thiazole.

Alternatively, of the methods of using compounds of formula (I) to treatcancer, inflammation or insulin resistance as set forth above in theSummary of the Invention, a preferred group of methods is that groupwherein the compound of formula (I) is a compound of formula (I) whereinA is an optionally substituted straight or branched alkylene chain offour carbons, or an optionally substituted straight or branchedalkenylene chain of four carbons.

Another preferred group of methods is that group wherein the compound offormula (I) is a compound of formula (I) wherein A is —R⁸—C(R²)—N(R⁵)—or —R⁸—N(R⁵)—C(R²)—.

Another preferred group of methods is that group wherein the compound offormula (I) is a compound of formula (I) wherein A is —R⁸—O—C(R²)—,—R⁷—O—C(R²)—R⁷—, or —R⁹—C(R²)—.

Another preferred group of methods is that group wherein the compound offormula (I) is a compound of formula (I) wherein A is —R⁸—O—R⁷—,—R⁸—S(O)_(t)—R⁷— (where t is 0 to 2), or —R⁹—O—.

Another preferred group of methods is that group wherein the compound offormula (I) is a compound of formula (I) wherein A is—R⁷—C(R²)—N(R⁵)S(O)_(t)— (where t is 0 to 2) or.

Another preferred group of methods is that group wherein the compound offormula (I) is a compound of formula (I) wherein A is —R⁸—N(R⁵)—R⁷—.

Another preferred group of methods is that group wherein the compound offormula (I) is a compound of formula (I) wherein A is—R⁸—C(R²)—N(R⁵)—S(O)_(t)NH—R¹⁴.

Of the methods and the preferred group of methods set forth above, apreferred subgroup of methods is that subgroup wherein the compound offormula (I) is a compound of formula (I) wherein R¹ is ═O.

Another preferred subgroup of methods is that subgroup wherein thecompound of formula (I) is a compound of formula (I) wherein R¹ is ═S.

Of the methods and the preferred groups and subgroups of methods setforth above, a preferred class of methods is that class wherein thecompound of formula (I) is a compound of formula (I) wherein R² is ═O.

Another preferred class of methods is that class wherein the compound offormula (I) is a compound of formula (I) wherein R² is ═S.

Of the methods and the preferred groups, subgroups and classes ofmethods as set forth above, a preferred subclass of methods is thatsubclass wherein the compound of formula (I) is a compound of formula(I) wherein R³ is hydrogen, alkyl, alkenyl, cycloalkyl, cycloalkylalkyl,or haloalkyl.

Another preferred subclass of methods is that subclass wherein thecompound of formula (I) is a compound of formula (I) wherein R³ is aryl,aralkyl, or aralkenyl.

Another preferred subclass of methods is that subclass wherein thecompound of formula (I) is a compound of formula (I) wherein R³ is halo,haloalkoxy, or —OR¹⁰.

Another preferred subclass of methods is that subclass wherein thecompound of formula (I) is a compound of formula (I) wherein R³ isnitro, cyano, or —N═N—O—R¹¹.

Another preferred subclass of methods is that subclass wherein thecompound of formula (I) is a compound of formula (I) wherein R³ is—C(O)OR¹⁰ or —C(O)N(R¹⁰)₂.

Another preferred subclass of methods is that subclass wherein thecompound of formula (I) is a compound of formula (I) wherein R³ is—N(R¹⁰)₂, —N(R¹⁰)C(O)R¹⁰, or —N(R¹⁰)C(O)OR¹¹.

Another preferred subclass of methods is that subclass wherein thecompound of formula (I) is a compound of formula (I) wherein R³ is—S(O)_(t)R¹⁰ (where t is 0 to 2) or —S(O)_(t)N(R¹⁰)₂ (where t is 0 to2).

Another preferred subclass of methods is that subclass wherein thecompound of formula (I) is a compound of formula (I) wherein R³ isheterocyclyl or heterocyclylalkyl.

Of the methods and the preferred groups, subgroups, classes andsubclasses of methods set forth above, a preferred set of methods isthat set wherein the compound of formula (I) is a compound of formula(I) wherein R⁴ is hydrogen, alkyl, haloalkyl, cycloalkyl, orcycloalkylalkyl.

Another preferred set of methods is that set wherein the compound offormula (I) is a compound of formula (I) wherein R⁴ is aralkyl or aryl.

Another preferred set of methods is that set wherein the compound offormula (I) is a compound of formula (I) wherein R⁴ is heterocyclyl orheterocyclylalkyl.

Of the methods and the preferred groups, subgroups, classes, subclassesand sets of methods as set forth above, a preferred subset of methods isthat subset wherein the compound of formula (I) is a compound of formula(I) wherein R⁶ is hydrogen, alkyl, alkenyl, cycloalkyl, cycloalkylalkyl,or haloalkyl.

Another preferred subset of methods is that subset wherein the compoundof formula (I) is a compound of formula (I) wherein R⁶ is aryl, aralkyl,or aralkenyl.

Another preferred subset of methods is that subset wherein the compoundof formula (I) is a compound of formula (I) wherein R⁶ is halo,haloalkoxy, or —OR¹⁰.

Another preferred subset of methods is that subset wherein the compoundof formula (I) is a compound of formula (I) wherein R⁶ is nitro, cyano,or —N═N—O—R¹¹.

Another preferred subset of methods is that subset wherein the compoundof formula (I) is a compound of formula (I) wherein R⁶ is —C(O)OR¹⁰ or—C(O)N(R¹⁰)₂.

Another preferred subset of methods is that subset wherein the compoundof formula (I) is a compound of formula (I) wherein R⁶ is —N(R¹⁰)₂,—N(R¹⁰)C(O)R¹⁰, or —N(R¹⁰)C(O)OR¹¹.

Another preferred subset of methods is that subset wherein the compoundof formula (I) is a compound of formula (I) wherein R⁶ is —S(O)_(t)R¹⁰(where t is 0 to 2) or —S(O)_(t)N(R¹⁰)₂ (where t is 0 to 2).

Another preferred subset of methods is that subset wherein the compoundof formula (I) is a compound of formula (I) wherein R⁶ is heterocyclylor heterocyclylalkyl.

Of the methods of using compounds of formula (Ia), a preferred subgroupof methods is that subgroup wherein the cancer is associated with thecolon, or colorectal cancer, or cancer associated withhyperproliferation or tissue remodelling or repair. Of this preferredsubgroup, a preferred class of methods is that class wherein the canceris associated the activity of PTPN12 or PTPN2.

Of this preferred group, subgroup and class, a preferred subclass ofmethods is that subclass wherein the compound of formula (Ia) is acompound of formula (Ia) wherein:

A is —R^(8a)—C(R^(2a))—N(R^(5a))—;

R^(1a) and R^(2a) are each independently ═O or ═S;

R^(3a) is alkyl, aryl, aralkyl, chloro, iodo, bromo, haloalkyl,haloalkoxy, nitro, cyano, —N═N—O—R^(11a), —OR^(12a), —C(O)OR^(10a),—C(O)N(R^(10a))₂—N(R^(12a))₂, —N(R^(10a))C(O)R^(10a),—N(R^(10a))C(O)OR^(11a), —S(O)_(t)R^(10a) (where t is 0 to 2), or—S(O)_(t)N(R^(10a))₂ (where t is 0 to 2);

R^(4a) is hydrogen or alkyl;

R^(5a) is hydrogen or alkyl;

R^(2a) is alkyl, aryl, aralkyl, halo, haloalkyl, haloalkoxy, nitro,cyano, —N═N—O—R^(11a), —C(O)N(R^(10a))₂, —N(R^(10a))₂,—N(R^(10a))C(O)R^(10a), —N(R^(10a))C(O)OR^(11a), —S(O)_(t)R^(10a) (wheret is 0 to 2), —S(O)_(t)N(R^(10a))₂ (where t is 0 to 2), or—S(O)_(t)NH—R^(14a);

each R^(8a) is an optionally substituted straight or branched alkyleneor alkenylene chain of two carbons;

each R^(10a) is hydrogen, alkyl, aralkyl or aryl;

R^(11a) is hydrogen, alkyl or aralkyl;

R^(12a) is hydrogen, aryl or aralkyl; and

R^(14a) is a thiazole.

Of this preferred subclass of methods, a preferred set of methods isthat set wherein the compound of formula (Ia) is a compound of formula(Ia) wherein:

A is —R^(8a)—C(R^(2a))—N(R^(5a))—;

R^(1a) and R^(2a) are each independently ═O or ═S;

R^(3a) is alkyl, aryl, aralkyl, chloro, iodo, bromo, haloalkyl, orhaloalkoxy;

R^(3a) is hydrogen or alkyl;

R^(5a) is hydrogen or alkyl;

R^(6a) is —C(O)N(R^(10a))₂, —S(O)_(t)R^(10a) (where t is 0 to 2),—S(O)_(t)N(R^(10a))₂ (where t is 0 to 2), or —S(O)_(t)NH—R^(14a); where

each R^(8a) is an optionally substituted straight or branched alkyleneor alkenylene chain of two carbons; and

each R^(10a) is hydrogen, alkyl, aralkyl or aryl; and

R^(14a) is a thiazole.

Of this preferred set of methods, a preferred subset of methods is thatsubset wherein the compound of formula (Ia) is a compound of formula(Ia) wherein:

A is —R^(8a)—C(R^(2a))—N(R^(5a))—;

R^(1a) and R^(2a) are both ═O;

R^(3a) is alkyl, haloalkyl or haloalkoxy;

R⁴′ is hydrogen;

R^(5a) is hydrogen or alkyl;

R^(6a) is —S(O)₂N(R^(10a))₂;

R^(8a) is ethylene; and

each R^(10a) is hydrogen or alkyl.

Alternatively, of the methods of using compounds of formula (Ia) totreat inflammation in a mammal as set forth above in the Summary of theInvention, a preferred group of methods is that group wherein thecompound of formula (Ia) is a compound of formula (Ia) wherein A is anoptionally substituted straight or branched alkylene chain of fourcarbons or an optionally substituted straight or branched alkenylenechain of four carbons.

Another preferred group is that group of methods wherein the compound offormula (Ia) is a compound of formula (Ia) wherein A is—R^(8a)—C(R^(2a))—N(R^(5a))— or —R^(8a)—N(R^(5a))—C(R^(2a))—.

Another preferred group is that group of methods wherein the compound offormula (Ia) is a compound of formula (Ia) wherein A is—R^(8a)—O—C(R^(2a))—, —R^(7a)—O—C(R^(2a))—R^(7a)—, or R^(9a)—C(R^(2a))—.

Another preferred group is that group of methods wherein the compound offormula (Ia) is a compound of formula (Ia) wherein A is—R^(8a)-O-R^(7a)—, —R^(8a)—S(O)_(t)—R^(7a)— (where t is 0 to 2), or—R^(9a)—O—.

Another preferred group is that group of methods wherein the compound offormula (Ia) is a compound of formula (Ia) wherein A is—R^(7a)—C(R^(2a))—N(R^(5a))—S(O)_(t)— (where t is 0 to 2).

Another preferred group is that group of methods wherein the compound offormula (Ia) is a compound of formula (Ia) wherein A is—R^(8a)—N(R^(5a))—R^(7a)— or —R^(9a)—N(R^(5a))—.

Of the methods and the preferred groups of methods set forth above, apreferred subgroup of methods is that subgroup wherein the compound offormula (Ia) is a compound of formula (Ia) wherein R^(1a) is ═O.

Another preferred subgroup is that subgroup of methods wherein thecompound of formula (Ia) is a compound of formula (Ia) wherein R^(1a) is═S.

Of the methods and preferred groups and subgroups of methods set forthabove, a preferred class of methods is that class wherein the compoundof formula (Ia) is a compound of formula (Ia) wherein R^(2a) is ═O.

Another preferred class is that class of methods wherein the compound offormula (Ia) is a compound of formula (Ia) wherein R^(2a) is ═S.

Of the methods and the preferred groups, subgroups and classes ofmethods set forth above, a preferred subclass of methods is thatsubclass wherein the compound of formula (Ia) is a compound of formula(Ia) wherein R^(3a) is alkyl, alkenyl, cycloalkyl, cycloalkylalkyl orhaloalkyl.

Another preferred subclass is that subclass of methods wherein thecompound of formula (Ia) is a compound of formula (Ia) wherein R^(3a) ischloro, iodo, bromo, haloalkoxy or —OR^(10a).

Another preferred subclass is that subclass of methods wherein thecompound of formula (Ia) is a compound of formula (Ia) wherein R^(3a) isnitro, cyano, or —N═N—O—R^(11a).

Another preferred subclass is that subclass of methods wherein thecompound of formula (Ia) is a compound of formula (Ia) wherein R^(3a) is—C(O)OR^(10a) or —C(O)N(R^(10a))₂.

Another preferred subclass is that subclass of methods wherein thecompound of formula (Ia) is a compound of formula (Ia) wherein R^(3a) is—N(R^(12a))₂, —N(R^(10a))C(O)R^(10a) or —N(R^(10a))C(O)OR^(11a).

Another preferred subclass is that subclass of methods wherein thecompound of formula (Ia) is a compound of formula (Ia) wherein R^(3a) is—S(O)_(t)R¹⁰ (where t is 0 to 2) or —S(O)_(t)N(R^(10a))₂ (where t is 0to 2).

Another preferred subclass is that subclass of methods wherein thecompound of formula (Ia) is a compound of formula (Ia) wherein R^(3a)heterocyclylalkyl.

Of the methods and the preferred groups, subgroups, classes andsubclasses of methods set forth above, a preferred set is that set ofmethods wherein the compound of formula (Ia) is a compound of formula(Ia) wherein R^(4a) is hydrogen, alkyl, haloalkyl, cycloakyl, orcycloalkyl.

Another preferred set is that set of methods wherein the compound offormula (Ia) is a compound of formula (Ia) wherein R^(4a) is aralkyl oraryl.

Another preferred set is that set of methods wherein the compound offormula (Ia) is a compound of formula (la) wherein R^(4a) isheterocyclyl or heterocyclylalkyl.

Of the methods and the groups, subgroups, classes, subclasses and setsof methods set forth above, a preferred subset is that subset of methodswherein the compound of formula (Ia) is a compound of formula (Ia)wherein R^(6a) is alkyl, alkenyl, cycloalkyl, cycloalkylalkyl orhaloalkyl.

Another preferred subset is that subset of methods wherein the compoundof formula (Ia) is a compound of formula (Ia) wherein R^(6a) is halo orhaloalkoxy.

Another preferred subset is that subset of methods wherein the compoundof formula (Ia) is a compound of formula (Ia) wherein R^(6a) is nitro,cyano, or —N═N—O—R^(11a).

Another preferred subset is that subset of methods wherein the compoundof formula (Ia) is a compound of formula (Ia) wherein R^(6a) is—C(O)N(R^(10a))₂.

Another preferred subset is that subset of methods wherein the compoundof formula (Ia) is a compound of formula (Ia) wherein R^(6a) is—N(R^(10a))₂, —N(R^(10a))C(O)R^(10a) or —N(R^(10a))C(O)OR^(11a).

Another preferred subset is that subset of methods wherein the compoundof formula (Ia) is a compound of formula (Ia) wherein R^(6a) is—S(O)_(t)R¹⁰ (where t is 0 to 2) or —S(O)_(t)N(R^(10a))₂ (where t is 0to 2).

Another preferred subset is that subset of methods wherein the compoundof formula (Ia) is a compound of formula (Ia) wherein R^(6a)heterocyclylalkyl or heterocyclylalkyl.

Of the pharmaceutical compositions comprising a pharmaceuticallyacceptable excipient and a compound of formula (Ia) as set forth abovein the Summary of the Invention, a preferred group of pharmaceuticalcompositions is that group wherein the compound of formula (Ia) is acompound of formula (Ia) wherein A is an optionally substituted straightor branched alkylene chain of four carbons or an optionally substitutedstraight or branched alkenylene chain of four carbons.

Another preferred group is that group of pharmaceutical compositionswherein the compound of formula (Ia) is a compound of formula (Ia)wherein A is —R^(8a)—OC(R^(2a))—N(R^(5a))- or—R^(8a)—N(R^(5a))—C(R^(2a))—.

Another preferred group is that group of pharmaceutical compositionswherein the compound of formula (Ia) is a compound of formula (Ia)wherein A is —R^(8a)—O—C(R^(2a)), —R^(7a)-O—C(R^(2a))—R^(7a)— or—R^(9a)—C(R^(2a))—.

Another preferred group is that group of pharmaceutical compositionswherein the compound of formula (Ia) is a compound of formula (Ia)wherein A is —R^(8a)—O—R^(7a)—, —R^(8a)—S(O)_(t)—R^(7a)— (where t is 0to 2), or —R^(9a)—O—.

Another preferred group is that group of pharmaceutical compositionswherein the compound of formula (Ia) is a compound of formula (Ia)wherein A is —R^(7a)—C(R^(2a))—N(R^(5a))—S(O)_(t)— (where t is 0 to 2).

Another preferred group is that group of pharmaceutical compositionswherein the compound of formula (Ia) is a compound of formula (Ia)wherein A is —R^(8a)—N(R^(5a))—R^(7a)— or R^(9a)—N(R^(5a))—.

Of the pharmaceutical compositions and the preferred groups ofpharmaceutical compositions set forth above, a preferred subgroup ofpharmaceutical compositions is that subgroup wherein the compound offormula (Ia) is a compound of formula (Ia) wherein R^(1a) is ═O.

Another preferred subgroup is that subgroup of pharmaceuticalcompositions wherein the compound of formula (Ia) is a compound offormula (Ia) wherein R^(1a) is ═S.

Of the pharmaceutical compositions and the preferred groups andsubgroups of pharmaceutical compositions set forth above, a preferredclass of pharmaceutical compositions is that class wherein the compoundof formula (Ia) is a compound of formula (Ia) wherein R^(2a) is ═O.

Another preferred class is that class of pharmaceutical compositionswherein the compound of formula (Ia) is a compound of formula (Ia)wherein R^(2a) is ═S.

Of the pharmaceutical compositions and the preferred groups, subgroupsand classes of pharmaceutical compositions set forth above, a preferredsubclass of pharmaceutical compositions is that subclass wherein thecompound of formula (Ia) is a compound of formula (Ia) wherein R^(3a) isalkyl, alkenyl, cycloalkyl, cycloalkylalkyl or haloalkyl.

Another preferred subclass is that subclass of pharmaceuticalcompositions wherein the compound of formula (Ia) is a compound offormula (Ia) wherein R^(3a) is chloro, iodo, bromo, haloalkoxy or—OR^(10a).

Another preferred subclass is that subclass of pharmaceuticalcompositions wherein the compound of formula (Ia) is a compound offormula (Ia) wherein R^(3a) is nitro, cyano, or —N═N—O—R^(11a).

Another preferred subclass is that subclass of pharmaceuticalcompositions wherein the compound of formula (Ia) is a compound offormula (Ia) wherein R^(3a) is —C(O)OR^(10a) or —C(O)N(R^(10a))₂.

Another preferred subclass is that subclass of pharmaceuticalcompositions wherein the compound of formula (Ia) is a compound offormula (Ia) wherein R^(3a) is —N(R^(12a))₂, N(R^(10a))C(O)R^(10a) or—N(R^(10a))C(O)OR^(11a).

Another preferred subclass is that subclass of pharmaceuticalcompositions wherein the compound of formula (Ia) is a compound offormula (Ia) wherein R^(3a) is —S(O)_(t)R¹⁰ (where t is 0 to 2) or—S(O)_(t)N(R^(10a))₂ (where t is 0 to 2).

Another preferred subclass is that subclass of pharmaceuticalcompositions wherein the compound of formula (Ia) is a compound offormula (Ia) wherein R^(3a) heterocyclylalkyl.

Of the pharmaceutical compositions and the preferred groups, subgroups,classes and subclasses of pharmaceutical compositions set forth above, apreferred set is that set of pharmaceutical compositions wherein thecompound of formula (Ia) is a compound of formula (Ia) wherein R^(4a) ishydrogen, alkyl, haloalkyl, cycloakyl, or cycloalkyl.

Another preferred set is that set of pharmaceutical compositions whereinthe compound of formula (Ia) is a compound of formula (Ia) whereinR^(4a) is aralkyl or aryl.

Another preferred set is that set of pharmaceutical compositions whereinthe compound of formula (Ia) is a compound of formula (Ia) whereinR^(4a) is heterocyclyl or heterocyclylalkyl.

Of the pharmaceutical compositions and the groups, subgroups, classes,subclasses and sets of pharmaceutical compositions set forth above, apreferred subset is that subset of pharmaceutical compositions whereinthe compound of formula (Ia) is a compound of formula (Ia) whereinR^(6a) is alkyl, alkenyl, cycloalkyl, cycloalkylalkyl or haloalkyl.

Another preferred subset is that subset of pharmaceutical compositionswherein the compound of formula (Ia) is a compound of formula (Ia)wherein R^(6a) is halo or haloalkoxy.

Another preferred subset is that subset of pharmaceutical compositionswherein the compound of formula (Ia) is a compound of formula (Ia)wherein R^(6a) is nitro, cyano, or —N═N—O—R^(1a).

Another preferred subset is that subset of pharmaceutical compositionswherein the compound of formula (Ia) is a compound of formula (Ia)wherein R^(6a) is —C(O)N(R^(10a))₂.

Another preferred subset is that subset of pharmaceutical compositionswherein the compound of formula (Ia) is a compound of formula (Ia)wherein R^(6a) is —N(R^(10a))₂, —N(R^(10a))C(O)R^(10a) orN(R^(10a))C(O)OR^(11a).

Another preferred subset is that subset of pharmaceutical compositionswherein the compound of formula (Ia) is a compound of formula (Ia)wherein R^(6a) is —S(O)_(t)R¹⁰ (where t is 0 to 2) or—S(O)_(t)N(R^(10a))₂ (where t is 0 to 2).

Another preferred subset is that subset of pharmaceutical compositionswherein the compound of formula (Ia) is a compound of formula (Ia)wherein R^(6a) is heterocyclyl or heterocyclylalkyl.

Of the methods of treating a mammalian cell with a compound of formula(I) wherein the method comprises administering the compound of formula(I) to a mammalian cell and the compound of formula (I) is capable ofinhibiting the activity of PTPN12, PTPN2 and/or PTPN1, a preferred groupof methods is that group wherein the mammalian cell is treated in vitro.

Another preferred group of these methods is that group wherein themammalian cell is treated in vivo.

Another preferred group of these methods is that group wherein theinhibition of activity results in a reduction of cell adhesion, celldivision, cell migration, or tumor growth.

Another preferred group of these methods is that group wherein theinhibition of activity results in control of lymphocyte activation.

Another preferred group of these methods is that group wherein theinhibition of PTPN1 activity results in control of insulin resistance.

Preparation of the Compounds of Formula (I) and Formula (Ia)

Compounds of formula (I) and formula (Ia) in the methods andpharmaceutical compositions of the invention may be prepared accordingto methods known to one skilled in the art, or by the methods similar tothose disclosed in Gorishnii, V. Ya. et al., Farm. Zh. (Kiev) (2001),Vol. 2, pp. 64-67; Gorishnyi, V. Ya. et al., Farm. Zh. (Kiev) (1995),Vol. 4, pp. 50-53; PCT Published Patent Application WO 00/76988;European Patent Specification 0 047109; U.S. Pat. No. 5,310,618; or U.S.Pat. No. 4,965,155; or by the following method.

It is understood that in the following description, combinations ofsubstituents and/or variables of the depicted formulae are permissibleonly if such contributions result in stable compounds.

It will also be appreciated by those skilled in the art that in theprocess described below the functional groups of intermediate compoundsmay need to be protected by suitable protecting groups. Such functionalgroups include hydroxy, amino, mercapto and carboxylic acid. Suitableprotecting groups for hydroxy include trialkylsilyl or diarylalkylsilyl(e.g., t-butyldimethylsilyl, t-butyidiphenylsilyl or trimethylsilyl),tetrahydropyranyl, benzyl, and the like. Suitable protecting groups foramino, amidino and guanidino include t-butoxycarbonyl,benzyloxycarbonyl, and the like. Suitable protecting groups for mercaptoinclude —C(O)—R (where R is alkyl, aryl or aralkyl), p-methoxybenzyl,trityl and the like. Suitable protecting groups for carboxylic acidinclude alkyl, aryl or aralkyl esters.

Protecting groups may be added or removed in accordance with standardtechniques, which are well-known to those skilled in the art and asdescribed herein.

The use of protecting groups is described in detail in Green, T. W. andP. G. M. Wutz, Protective Groups in Organic Synthesis (1991), 2nd Ed.,Wiley-Interscience. The protecting group may also be a polymer resinsuch as a Wang resin or a 2-chlorotrityl chloride resin.

It will also be appreciated by those skilled in the art, although suchprotected derivatives of compounds of formulae (I), as described abovein the Summary of the Invention, may not possess pharmacologicalactivity as such, they may be administered to a mammal with cancer orinflammation and thereafter metabolized in the body to form compounds ofthe invention which are pharmacologically active. Such derivatives maytherefore be described as “prodrugs”. All prodrugs of compounds offormula (I) and formula (Ia) are included within the scope of theinvention.

The following Reaction Scheme depicts the preparation of compounds offormula (I) where p, A, R¹, R³, R⁴ and R⁶ are described above in theSummary of the Invention, and X is halo and R¹³ is hydrogen:

In this general scheme, starting components may be obtained from sourcessuch as Aldrich, or synthesized according to sources known to those ofordinary skill in the art, e.g., Smith and March, March's AdvancedOrganic Chemistry: Reactions, Mechanisms, and Structure, 5th edition(Wiley Interscience, New York).

In general, dithiocarbamate compounds of formula (C) may be preparedaccording to Step 1 of the reaction scheme depicted herein, wherebycarbon disulfide (i.e., the compound of formula (A)) at a concentrationof about 3.5 moles/liter is reacted with about an equimolar quantity ofan amine compound of formula (B), in the presence of ammonium hydroxidesolution at about 0° C. The admixture is warmed to ambient temperature,stirred for up to about 18 hours, and concentrated to dryness. Theresulting substance is a compound of formula (C).

Rhodanine-derived compounds of formula (E) can be prepared undercyclization conditions according to schemes known to those of ordinaryskill in the art. For instance, a compound of formula (E) is formedaccording to Step 2 of the reaction scheme depicted herein, whereby theforegoing quantity of the compound of formula (C) is reacted with aboutan equimolar quantity of a compound of formula (D) (wherein R¹ is O orS) or a basic salt thereof, in an aqueous solution (at about 0° C.)alkalized with dilute sodium carbonate. The reaction mixture is warmedto ambient temperature, admixed with about 6.4 volumes of warm 5 Mhydrochloric acid (about 70° C.), and heated to about 90° C. for about 1hour. After cooling, the resulting precipitate is isolated byfiltration, washed with water and allowed to dry, affording a compoundof formula (E).

Compounds of formula (F) can be obtained from sources such as Aldrich,or prepared according to schemes known to those of ordinary skill in theart. In one aspect, nitro-substituted benzaldehyde compounds may beprepared under standard aromatic substitution conditions, such as bytreatment of benzaldehyde with nitric acid and sulfuric acid. In anotheraspect, halogen-substituted benzaldehyde compounds may be prepared understandard aromatic substitution conditions, such as by treatment ofbenzaldehyde with naturally-occurring diatomic halogen compounds (i.e.,F₂, Cl₂, Br₂, or I₂) with iron metal. In yet another aspect,alkyl-substituted benzaldehyde compounds may be prepared under standardaromatic substitution conditions, such as by Friedel-Crafts alkylationof benzaldehyde with an alkyl halide in the presence of an aluminumhalide compound. Such treatments normally produce mixtures comprisingcompounds with substitutions in various different ring positions, thoughspecific chemical properties of the reagents used, particularly thearomatic compound, may promote the synthesis of certain compounds withspecific substitutions at desired ring positions as major synthesisproducts. Collection of pure major and/or minor synthesis products maybe achieved with the use of a preparative separation and isolationtechnique such as high performance liquid chromatography (HPLC).

Compounds of formula (G) can be prepared under standard condensationreaction conditions according to schemes known to those of ordinaryskill in the art. For instance, a compound of formula (G) is formedaccording to Step 3 of the reaction scheme depicted herein, whereby acompound of formula (E) is combined with about an equimolar quantity ofa substituted benzene compound of formula (F) (wherein R³ and R⁴ areselected from constituents as defined in the Summary of the Invention)in an aqueous solution containing sodium acetate and acetic acid. Thereaction is heated to reflux for up to 16 hours with stirring. Aftercooling, the resulting precipitate is isolated by filtration, washedwith water and allowed to dry, affording a compound of formula (G), as astereoisomer, stereoisomer mixture or as a pharmaceutically acceptablesalt thereof.

Compounds of formula (G) wherein R¹¹ is hydrogen is reacted with acompound of formula (H) under standard amine alkylation conditions (asdepicted in Step 4 above) to afford a compound of formula (I). In aseparate optional aspect, a compound of formula (C) wherein R¹¹ ishydrogen may be reacted with a compound of formula (H) under standardamine alkylation conditions, the product of which can then proceedsequentially through Steps 2 and 3 of the reaction scheme as depictedherein, to afford a compound of formula (I). In another separateoptional aspect, a compound of formula (E) wherein R¹¹ is hydrogen maybe reacted with a compound of formula (H) under standard aminealkylation conditions, the product of which can then proceed throughStep 3 of the reaction scheme as depicted herein, to afford a compoundof formula (I).

Compounds of formula (Ia) as defined above in the Summary of theInvention may be prepared in a similar manner as described above.

The following examples are put forth so as to provide those of ordinaryskill in the art with a complete disclosure and description of how tomake and use the subject invention, and are not intended to limit thescope of what is regarded as the invention. Efforts have been made toensure accuracy with respect to the numbers used (e.g. amounts,temperature, concentrations, etc.) but some experimental errors anddeviations should be allowed for. Unless otherwise indicated, parts areparts by weight, molecular weight is average molecular weight,temperature is in degrees centigrade; and pressure is at or nearatmospheric.

EXAMPLE 1 Enzyme Preparation and Use

A. PTPN2

PTPN2 was cloned from a human placental cDNA library in the IMPACT™ (NewEngland BioLabs) bacterial expression system. The technology was firstdescribed by Chong et al., Gene (1997), Vol. 192, pp. 271-281. TheIMPACT™ Protein Purification System was purchased commercially from NewEngland BioLabs. The resulting product was used in the development of aprotein phosphatase assay for high-throughput screening (HTS) of targetmolecules and in other assays described herein (see Example 2).

Biochemical analysis performed on recombinant human PTPN2 fusion proteinexhibited protein phosphatase activity in the order of 1500 to 2500pmol/min/μg measured as phosphate release from a synthetic tyrosinephosphorylated peptide. This activity was considered to be in the highrange as compared to other recombinant protein tyrosine phosphatasesassayed. PTPN2 preparations were subsequently used extensively in invitro assays for the initial discovery of compounds having the abilityto inhibit PTPN2 activity.

B. PTPN12

PTPN12 was cloned in the IMPACT™ (New England BioLabs) bacterialexpression system. The IMPACT™ Protein Purification System was purchasedcommercially from New England BioLabs.

1. Cloning of Truncated Human PTPN12 into pTWIN-II Expression Vector

Expression of human truncated PTPN12 (PTP-PEST-N) as a fusion proteinrequired that the cDNA be ligated into the polyclonal site situated inframe and upstream of the intein gene of the IMPACT™ expression vectorpTWIN-II. The truncated version was used as it was far easier to handleand gave parallel results to the full length protein in comparisontesting. For the purpose of simplicity, PTP-PEST-N will be usedinterchangeably with PTPN12 in these Examples.

The PTPN12 coding sequence was generated by polymerase chain reaction(PCR) using gene-specific primers.

2. Human PTPN12 Expression and Purification

Active PTPN12 enzyme is expressed from the IMPACT™ vector system in thebacterial strain ER2566. Recombinant PTPN12 protein is purified frombacterial cells using affinity chromatography on chitin-agarose beadsfollowed by a chemical process whereby PTPN12 is released from itsaffinity tag. A complete quantitative and qualitative analysis of theprotein is monitored using Coomassie-Blue staining of SDS-PAGE separatedpreparations and by PTPN12-specific western blotting. PTPN12 is producedat levels in the range of 0.1-0.5 mg per litre of bacterial cellculture.

3. PTPN12 In vitro Phosphatase Assay

Biochemical analysis is performed on recombinant human PTPN12 fusionprotein. Typically, the PTPN12 preparations are found to exhibit proteinphosphatase reactivity in the order of 1500 to 2500 pmol/min/μg measuredas phosphate release from a synthetic tyrosine phosphorylated peptide.This activity is considered to be in the high range as compared to otherrecombinant protein tyrosine phosphatases. PTPN12 preparations weresubsequently used extensively in in vitro assays for the initialdiscovery of compounds having the ability to inhibit PTPN12 activity.

EXAMPLE 2 In Vitro Activity Profile for Phosphatases

Compounds of formula (I) and formula (Ia) were tested in the followingassay for their ability to inhibit the activity of the desiredphosphatase.

A. Reagent Preparation:

1. Malachite Green-Ammonium Molybdate Reagent

Two solutions were first prepared. Solution 1 contained 4.2% ammoniummolybdate tetrahydrate (Sigma, Cat# A-7302) in 4 N HCl. Solution 2contained 0.045% Malachite Green (Sigma, Cat. # M-9636). The twosolutions were mixed as follows: 250 mL of solution 1 and 750 mLsolution 2 with constant stirring for 20 min. The resulting mixture wasfiltered through 0.22 μM filter (one can use Nalgene™ bottle top vacuumfilters Cat # 28199-317). The solution was stored in a brown bottle at4° C.

B. Preparation of 1 mM ppC SRC 60 Substrate

The peptide sequence: TSTEPQY(PO4)QPGENL was prepared by conventionalmethods. Of this, 154 mg was dissolved in 100 mL dH₂O and the solutionvortexed until the peptide dissolved completely. The ppC SRC 60 was thenstored in 1 mL aliquots at −20° C. This is the “Substrate” used forpreparing the substrate working stock solution.

C. Procedure for Assay

The enzyme (phosphatase) activity was determined in a reaction thatmeasured phosphate relase from tyrosine phospho-specific peptides usinga method first described by Harder et al., Biochem. J. (1994), Vol. 298,pp. 395-401. This is a non-radioactive method for measuring freephosphate by the malachite green method first described by Van Veldhovenand Mannaerts, Anal Biochem. (1987), Vol. 161, pp. 45-48. 10× assaybuffer (250 mM Tris:100 mM, β-mercaptoethanol, 50 mM EDTA; pH 7.2) wasdiluted to 5× concentration (conc.) with distilled H₂O (dH₂O). Then 71.4μM of substrate working stock solution was prepared in dH₂O.

In a microcentrifuge tube, the required volume. of enzyme stock waspipetted, diluted with the required volume of 5× assay buffer and mixed.

The colour reagent was prepared by thoroughly mixing 10 mL malachitegreen-ammonium molybdate reagent and 100 μL of 1% Tween-20 (1 mLTween-20 (BDH, #06435) dissolved in 99 mL dH₂O) into a reagent reservoirand stored at room temperature. Approximately 10 mL of colour reagent isrequired per assay plate, or 100 μL per well.

Sample Compound Preparation

In a Falcon 96 well plate the sample compound was diluted in 1% DMSO (1mL DMSO (Sigma, Cat. # D-8779) dissolved in 99 mL dH₂O and stored atroom temperature) such that the concentration of the sample compoundworking stock solution is ten times the final desired concentration ofthe compound in the assay.

The working stock solution was prepared as per the requiredconcentration of sample compound in the assay.

The negative control consisted of 5 μl 1% DMSO and 35 μL substrateworking stock solution and 10 μL diluted enzyme, per well, and wasplaced in the first column of wells on the plate. The last column ofwells on the plate was reserved for an enzyme blank, which consisted of5 μL 1% DMSO, 35 μL substrate working stock solution, and 10 μL 5× assaybuffer, per well. Test samples were placed in columns 2-11 and consistedof 5 μL sample in 1% DMSO, 35 μL substrate working stock solution, and10 μL of diluted enzyme, per well, at the desired concentration. Usingthe repeater function of a Biohit Multichannel™ pipettor, 5 μL of 100 μMsample from the Falcon™ plate columns was added to corresponding Costar™assay plate columns.

Then 5 μL 1% DMSO was added to column 1 & 12, and 10 μL of 5× assaybuffer to column 12.

Using a multichannel pipettor 35 μL of 71.4 μM ppC-SRC 60 substrate wasadded to all assay wells, then 10 μL of appropriately diluted enzyme wasadded to the wells on a column by column basis, pausing 5 secondsbetween columns. Timing started at the first addition.

The assay plate was incubated at room temperature (21° C.) for 15minutes. The reaction was “stopped” by adding 100 μL color reagent on acolumn by column basis, pausing 5 seconds between columns. Color wasallowed to develop for at least 15 minutes, but no longer than twohours, at room temperature. The plate was “read” on Bio-tek InstrumentsEL312e microplate Bio-Kinetics™ Reader at 590 nm and the data collectedas per instrument manual.

Data analysis was performed as follows. The blank and negative controlswere read, and blanks were subtracted from the average of negativecontrol values and sample values, and the % inhibition was expressed bythe following formula:% Inhibition=100−[corrected sample reading/corrected Negative Controlreading*100].

Compounds of the invention showed the following profile of inhibition:TABLE 1 Inhibition Inhibition Inhibition of PTPN12 of PTPN2 of PTPN1Compound IC₅₀ IC₅₀ IC₅₀ 3-[5-(4-methylbenzylidene)-4- 1.5 μm 1.6 3.0 μmoxo-2-thioxothiazolidin-3- yl]-N-(4-sulfamoylphenyl)- propionamide3-[5-(4-methylbenzylidene)-4- 1.1 μm 0.95 μm 1.5 μmoxo-2-thioxothiazolidin-3- yl]-N-[4-(thiazol-2- ylsulfamoyl)-phenyl]-propionamide

EXAMPLE 3 Cell Migration in a Boyden Chamber

A range of cell lines are used in this assay, particularly the prostatecancer cell line PC3 and PTPN12 mouse embryonic fibroblasts (MEFs). Therole of PTPN12 in migration was established based on the observations ofPTPN12 negative MEFs. Cell adhesion and migration are dynamic biologicalactivities involving the assembly and disassembly of a large number ofextracellular and intracellular molecules, for example, actin, which areregulated in turn by protein phosphorylation. Hence locking the systemin a phosphorylated (inhibition of phosphatases) or dephosphorylated(inhibition of kinases) state has a profound effect on theassembly/disassembly process and ultimately migration. Migration isreduced in PTPN12 knock-out MEFs. By extension, a PTPN12 inhibitorshould reduce cell migration in a Boyden chamber. Therefore, as areadout for PTPN12 activity, the following assay is designed to analyzecell migration in Boyden chambers. The Boyden assay is an experimentused to determine the capacity of a cell type to migrate onextracellular matrix. Unless otherwise indicated, all procedures areperformed under sterile conditions in a flow laminar hood and allincubations at 37° C. are performed in the CO₂ incubator.

A. Reagents

1. Staining Solution.

Calcein AM (Molecular Probes, Cat# C-1430) stain is prepared at 0.5ug/ml in Hanks Buffered Saline solution (GIBCO/BRL, Cat#14170-112).

2. Fibronectin Solution

A stock solution of fibronectin is prepared by dissolving 5 mg offibronectin: (Sigma, Cat: F-2006) in 5 mL of sterile phosphate-bufferedsolution (PBS) by up and down agitation with a P1000™ pipette. Theworking solution is prepared by mixing 100 μl of this stock solutionwith 10 mL of sterile PBS.

B. Assay (tumour cell lines)

For tumour cell lines, stock cells (ie. PC3 cells) are grown to 50-70%confluency in T175 flasks. Cells are trypsinized and a suspensionprepared to a concentration of 2×10⁵/ml in media without serum. To thetop chamber of each well of the HTS FluoroBlok™ 24-well insert systemplates (Cat# 351158) 450 μl of cell suspension (or media for controls)is added. Compounds for testing are prepared as 10× stocks in serum-freemedia from DMSO stocks, with a maximum final DMSO concentration of0.25%. 50 μl of compound (or DMSO control) is then added to each topchamber, while 750 μl of media containing 10% fetal bovine serum isadded to the bottom chamber as the chemoattractant. The plates areincubated for 20-24 hours at 37° C., 5% CO₂. Following incubation, theinsert plate is transferred into a second 24-well companion platecontaining 0.5 ml of 5 ug/ml calcein AM in HBSS and incubated for 1 hourat 37° C., 5% CO₂. Fluorescence of migrated cells is read in aFluoroskan Ascent FL™ (or equivalent) with bottom reading atexcitation/emission wavelength of 485/538 nm. Only those cells that havemigrated through the pores of the FluoroBlok™ membrane will be read. ForMEFs, the plates are coated on both sides of the membrane with 10 mg/mLfibronectin solution for 18 hours at 4° C. After incubation, the coatingsolution is removed by aspiration and the excess is washed twice withPBS. Cell seeding and detection are then performed as described fortumour cell lines.

C. Data Analysis

Data is expressed as fluorescence unit (FU) from the sum of middle 25areas per 24-well or as percentage of migration inhibition by followingformula: % of invasion inhibition=100-FU of compound treated cellinvasion/FU of DMSO treated cell invasion times 100. Background issubtracted from all values, with background being represented by themedia only controls. TABLE 2 % Inhibition of Compound Migration at 25 μM3-[5-(4-methylbenzylidene)-4-oxo-2- 55 thioxothiazolidin-3-yl]-N-(4-sulfamoylphenyl)-propionamide 3-[5-(4-methylbenzylidene)-4-oxo-2- 87thioxothiazolidin-3-yl]-N-[4-(thiazol-2-ylsulfamoyl)-phenyl]-propionamide

EXAMPLE 4 The Status of p130^(cas) Phosphorylation on Western Blots

Phosphotyrosine profiling of PTPN12-heterozygote and PTPN12-knockoutmouse fibroblasts showed that a protein migrating at 130 kDa isconstitutively hyperphosphorylated in the knockout cells (Côté, J. F.,et al., Biochemistry (1998), Vol. 37, No. 38, pp. 13128-13137). Thisprotein was identified as being p130′, a protein found in focal adhesioncomplexes. It also appeared that the hyperphosphorylation of p130^(cas)in the PTPN12 knockout cells resulted in defective cell motility andfocal adhesion turnover (Angers-Loustau et al., 1999).

This following assay measures p130^(cas) phosphorylation status as areadout of PTPN12 or other PTP activity such as PTP-1B. Briefly, thegeneral tyrosine phosphorylation state of all cellular proteins isreduced by incubating the cells in suspension and then plating the cellsonto fibronectin-coated plates, thereby stimulating tyrosinephosphorylation through the integrin pathway. Following cell lysis,p130^(cas) immunoprecipitation and Western blotting using 4G10antiphosphotyrosine antibody are used to measure the tyrosinephosphorylation status of p130^(cas). A low level of p130^(cas) tyrosinephosphorylation is indicative of a high PTPN12 activity. The assay isperformed using PTPN12 knockout and heterozygote mouse fibroblasts.

A. Materials

1. PTPN12+/− mouse fibroblasts (AC4+/−) and PTPN12−/− mouse fibroblasts(AC6−/−) as kindly provided by Michel Tremblay and colleagues from theCancer Centre at McGill University.

2. RIPA Buffer is made by mixing 50 mM Tris-HCl pH 7.2, 150 mM NaCl,0.1% SDS (BioShop, Cat#: SDS 001), 0.5% sodium deoxycholate 10% solution(Sigma, Cat: D-6750), 1% NP40 (BDH Laboratory Supplies, Cat: 56009 2L),1 mM sodium vanadate (Fisher Scientific, Cat: S454-50) 200 mM solution,and “complete protease inhibitor mixture” (Roche Cat. 1836153).

3. SDS sample buffer is prepared by mixing 62.5 mM Tris-HCl pH 6.8, 20%glycerol (BioShop, Cat#: Gly 001), 2% SDS, 5% β-mercaptoethanol (AcrosOrganics, Cat#: 12547-2500), and 0.025% bromophenol blue (EM Science,OmniPur™).

B. Fibronectin Stimulation

6-Well plates (Fisher Scientific, Cat: 08-772-1 B, Falcon No. 3530) arecoated for 18 hours at 4° C. with a 10 mg/mL fibronectin solution(Sigma, Cat: F-2006, Lot: 109H7602) (density of 1 g/cm²). A volume of950 μl of the fibronectin solution is added to each well. The plates arewashed 2 times by adding 2 mL of PBS at ambient temperature to each welland by removing the PBS by aspiration. PBS 1% BSA solution (2 mL) isadded to each well to block non-specific sites and the plates areincubated for 1 hour at 37° C. in CO₂ incubator. The blocking solutionis removed by aspiration and the wells are washed before adding thecells to the wells.

C. Addition of Cells

Before adding the cells (AC4+/− and AC6−/−) to the prepared plates, theyare washed and removed from 10 cm culture dishes by incubating them for10 minutes at 37° C. in the CO₂ incubator with 1.5 mL of Trypsin/EDTA(0.05% Trypsin, 0.53 mM EDTA) (GibcoBRL, Cat: 25300-054) solution.Detached cells are suspended in 5 mL of PBS at ambient temperature,placed in 15 mL conical tubes and centrifuged at 600 g on a clinicalcentrifuge for 5 minutes. PBS is removed by aspiration, then the cellsare counted using a hemacytometer and cell concentration is adjusted to1×10⁶ cells/mL in DMEM 0.5% BSA.

The cell suspension mixed with a test compound in an amount adequate toprovide a range of 25 to 50 μM concentration is incubated for 30 minutesat 37° C. in the CO₂ incubator with mixing every ten minutes. An aliquotis retained as a control to determine the basal phosphorylation levelbefore fibronectin-treatment. For fibronectin treatment, 3 mL of thecell suspension is added to the fibronectin matrix in order to obtain60% confluence (3×10⁶ cells/well) before incubating for 45 minutes at37° C. in CO₂ incubator. Each sample is performed in duplicate.

At the end of fibronectin stimulation or incubation in suspension, cellsare washed with ice-cold PBS supplemented with 1 mM sodiumorthovanadate. Cells are lysed directly on the plate by adding 0.5 mL ofice-cold RIPA buffer supplemented with protease inhibitors and 1 mMsodium vanadate. Plates are incubated at 4° C. with frequent agitationfor 10 minutes, then disrupted by repeated aspiration with a P1000™micropipette before transfer to 1.5 mL microcentrifuge tubes. Cellulardebris is pelleted at 13,000 rpm (10000 g) for 10 minutes at 4° C. in amicrocentrifuge, and supernatants are drawn off into fresh 1.5 mLmicrocentrifuge tubes

Protein concentration in the cell lysates is assayed using Bio-RadProtein concentration kit DC™ (Bio-Rad) according to manufacturer'sinstructions. Immunoprecipitation of p130^(cas) is performed with anamount of 250 mg protein adjusted in a final volume of 1 mL with RIPAbuffer supplemented with 1 mM vanadate and inhibitors.

For the immunoprecipitation, 1 mg (4 mL) of anti-p130^(cas) mousemonoclonal (Transduction Laboratories, Cat: P27820) is added to eachsample and the mixture is incubated for 2 hours at 4° C. on a rotatingdevice. As an immunoprecipitation control, the same amount of celllysate is incubated at this step with 1 mg (3 mL) of rabbit pre-immuneserum. Then 20 mL of resuspended Protein G-Agarose™ beads (GibcoBRL,Cat: 15920-010) is added and the mixture is incubated with agitation for1 hour at 4° C. on a rotating device. Immunoprecipitates are collectedby centrifugation at 2000 g for 5 minutes at 4° C. Pellets are washed 3times with 1 mL of ice-cold RIPA buffer (the supernatant is removed byaspiration). After final wash, the beads are resuspended into 60 mL ofSDS sample buffer.

D. SDS-PAGE and Western Blotting

30 μl of immunoprecipitate are separated on a 10% polyacrylamide gel for1.5 hours at 125V (p130^(cas) is a 130 kDa protein)

Briefly, nitrocellulose membranes are blocked with TBS-Tween (TBST): 20mM Tris-HCl, pH 7.2-7.4 (BioShop, Cat#: TRS 001)), 150 mM NaCl:(BioShop, Cat#: SOD 001) and 0.1% (v/v) Tween-20: (BioShop, Cat: TWN508)1% BSA for 1 hour with agitation at ambient temperature.Antiphosphotyrosine monoclonal antibody clone 4G10 (UpstateBiotechnologies) is used at a 1/1000 dilution in TBST 1% BSA andincubated for 1 hour with agitation at ambient temperature. Theanti-mouse-IgG-HRP (horseradish peroxidase) conjugate (JacksonLaboratories) is used at a 1/20000 dilution in TBST 1% BSA and incubatedfor 1 hour at ambient temperature.

E. Data Analysis

The data are analyzed as a function of p130^(cas) phosphorylationstatus.

Compounds of the invention tested demonstrate a higher level ofphosphorylation in the PTPN12−/− cells when compared to the PTPN12+/−cells after fibronectin-treatment. Inhibition of PTPN12 in the +/− cellsby a compound of the invention results in a higher phosphorylaton stateof p130^(cas) in the treated cells when compared to the non-treatedcells.

The foregoing assay is also used, with the appropriate starting reagentsand enzyme preparations, to test the ability of the compounds of theinvention to inhibit PTPN12 and PTPN1 activity.

EXAMPLE 5 Cell Proliferation

This procedure (Jelinkova, R. B. et al., “Antiproliferative effect of alectin- and anti-Thy-1.2 antibody-targeted HPMA copolymer-bounddoxorubicin on primary and metastatic human colorectal carcinoma and onhuman colorectal carcinoma transfected with the mouse Thy-1.2 gene”,Bioconjug. Chem. (2000, September-October), Vol. 11, No. 5, pp. 664-73)is used to assess the effect compounds have on various cell lines withrespect to proliferation. The rate of anchorage-independent growth ofvarious tumor cells is quantified by measuring the amount of freeisotopic thymidine that has been incorporated into the cells over aperiod of time. The effect of any compound to inhibit the proliferationof various tumor cells could be used as an indication of its ability toprevent disease progression in cancer.

Cultured tumour cells are harvested cells as per normal procedures: i.e.trypsinize, centrifuge and count cells. A volume of 90 μL is used toseed 5,000 cells/well in a 96 well plate. Cells are incubated for 24hours at 37° C. under 5% CO₂. After incubation, cells should be 80-90%confluent.

³H-thymidine (Amersham) is diluted in cell culture media to aconcentration of 100 μCi/mL. The test compound is diluted in thethymidine broth to 10× the final desired concentration.

Then 10 μL of diluted compound is added to the 90 μL of cells alreadypresent in the 96-well plates. Six replicates wells are done pertreatment in columns 2 to 11. Plates were mixed by rocking.

A known cytotoxic compound such as staurosporine is used in relativelyhigh concentrations as a positive control in column 1. Diluted DMSO isused as a negative control in column 12. The plate is incubated forexactly 24 hours at 37° C.

After incubation, plates are observed under the microscope for obviouscell death, abnormal cell shape, crystal formation of the compound, etc.Then 25 μL volume of cold 50% TCA is added slowly to the 100 μL volumealready in each well, and incubated for 1-2 hours at 4° C. The platesare then washed 5× in tap water and allowed to dry completely (usuallyovernight) at ambient temperature. Finally, 100 μL of scintillationfluid is added to each well and the plates are counted in a Wallac 1450Microbeta™ counter according to user manual instructions.

The amount of inhibition is determined by the following formula:% inhibition=100−[(AVG treatment−AVG positive control)/100(AVG negativecontrol−AVG positive control)] TABLE 3 % Inhibition of Proliferation at50 μM Compound H460 Cells PC3 Cells 3-[5-(4-methylbenzylidene)-4-oxo-2-45 35 thioxothiazolidin-3-yl]-N-(4-sulfamoylphenyl)- propionamide3-[5-(4-methylbenzylidene)-4-oxo-2- 43 47thioxothiazolidin-3-yl]-N-[4-(thiazol-2-ylsulfamoyl)-phenyl]-propionamide

EXAMPLE 6 Cytotoxicity Assay

This procedure is used to assess the effects compounds have on variouscell lines with respect to cell viability. Cell viability is quantifiedusing calcein AM(3′,6′-Di(O-acetyl)-2′,7′-bis[N,N-bis-(carboxymethyl)aminomethyl]-fluorescein,tetraacetoxymethyl ester) and measuring its conversion to a fluorescentproduct (calcein) with a fluorimeter.

The principle of this assay is based on the presence of ubiquitousintracellular esterase activity found in live cells. By enzymaticreaction of esterase, non-fluorescent cell-permeant calcein AM isconverted to the intensely fluorescent calcein. The polyanionic dyecalcein is retained within live cells, producing a green fluorescence inlive cells. It is a faster, safer, and better-correlated indicator ofcytotoxicity than alternative methods (e.g. 3H-Thymidine incorporation).Calcein AM is susceptible to hydrolysis when exposed to moisture,Therefore, prepare aqueous working solutions containing calcein AMimmediately prior to use, and used within about one day.

A kit available to do this assay is “LIVE/DEAD® Viability/CytotoxicityKit (L-3224)” by Molecular Probes.

Cells were collected from tissue culture flasks and trypsinized,centrifuged, resuspended and counted. Cells were seeded to obtain 80-90%confluence (for normal cells, 10,000 cells/well (8000 cells/well forHUVEC cells)). A cell concentration of 110,000 cells/mL (88,000cells/well for HUVEC cells) is prepared as 90 μL volume is used perwell.

Using an 8-channel multi-dispense pipettor, cells were seeded in thecentral rows of the plate (Nunclon™ 96 well flat-bottom plate), leavingthe peripheral top and bottom rows with same volume of media only. Theplates were incubated at 37° C., 5% CO₂ overnight for approximately 24hours.

For test compounds, cell culture media (e.g., RPMI+10% FBS), 10×compound solution of final desired concentration from 20 mM stockcompounds was prepared.

10 μl of this 10× compound solution is added to the 90 μL of cellsalready present in the 96 well plates and a known cytotoxic compoundfrom previous testing is used as a positive control. The negativecontrol is 100% DMSO diluted to the same factor as the compounds.

The plates are incubated at 37° C. for approximately 24 hours, and mediais aspirated after plates are spun at 2400 rpm for 10 min at ambienttemperature. 100 μL of 1× DPBS (without calcium chloride, withoutmagnesium chloride (GibcoBRL, cat#14190-144)) is added to each well.

The calcein AM solution is prepared by added 50 μg of calcein AM crystal(m.w.=994.87 g/mol, Molecular Probes) and anhydrous DMSO (Sigma Aldrich)to make 1 mM stock and diluting stock to 2)(the final desiredconcentration in 1× DPBS just before the assay. 100 μL of this 2× isadded to the 100 μL of DPBS in the wells and the plates are incubated atambient temperature for 30 minutes. Fluorescence data is read andrecorded (Fluoroskan Ascent® FL fluorimeter (excitation˜485 nm,emission˜527 nm)).

The values for replicates (usually six) are averaged and % inhibition iscalculated as follows:% inhibition=100−[(AVG treatment−AVG positive control)/(AVG negativecontrol−AVG positive control)*100] TABLE 4 % Cytotoxicity On NormalCells At 50 Mm HS27 Huvec LL-86 Compound cells cells cells3-[5-(4-methylbenzylidene)-4-oxo-2- 0 11 16thioxothiazolidin-3-yl]-N-(4- sulfamoylphenyl)-propionamide3-[5-(4-methylbenzylidene)-4-oxo-2- 0 42 0thioxothiazolidin-3-yl]-N-[4-(thiazol-2-ylsulfamoyl)-phenyl]-propionamide

EXAMPLE 7 In Vivo Tumour Efficacy Study

To test the efficacy of test compounds on H460 subcutaneous xenograftalone and in combination with doxorubicin.

Athymic nude female mice are used for this experiment. A group of 120mice are inoculated with five million H460 cells in 100 μL Matrigel™excipient (VWR Canada). Tumours are measured three times a week withdigital calipers and the tumour volumes calculated. When tumours havereached an average size of 100 mm³, about three weeks after tumourimplantation, the mice are randomized by tumour volume and divided intosix groups with 10 mice per group.

Treatments with test compounds continue for about 20 days, and will beoral (gavage), intravenous, subcutaneous, or intraperitoneal dependingon the known solubility of the test compound.

The study breakdown in tabular form: TABLE 5 2nd Dose Group TreatmentDose Route Schedule Treatment mg/kg Route Schedul

A PTE — — — None — — B Compound 200 Oral Daily for None — — mg/kg 20days C Vehicle — Oral Daily for Doxorubicin 5 IV Every

20 days days D Vehicle — Oral Daily for Doxorubicin 7 IV Every

20 days days E Compound 200 Oral Daily for Doxorubicin 5 IV Every

mg/kg 20 days days F Compound 200 Oral Daily for Doxorubicin 7 IV Every

mg/kg 20 days days

At study termination, the mice are anesthetized 3 hours after the lastdose of test compound, and plasma and tissues are harvested and frozen.Tumours are divided into the desired number of aliquots and fast frozenfor later analysis.

EXAMPLE 8 Cell Invasion in Matrigel™

This procedure is used to assess the compound effect on the tumor cellinvasion through Matrigel™-coated Fluoroblok™ inserts. Invasion allowstumor cells to spread to sites other that the primary tumor. BDBioscience's BioCoat FluoroBlok™ Invasion Systems™ combine the benefitsof the BD BioCoat Matrigel™ Invasion Chambers with the fluorescenceblocking membrane capabilities of the BD Falcon™ HTS FluoroBlok™24-Multiwell Insert System. The following assay uses this system toassess compound effects on the anti-tumor cell invasion through layer ofMatrigel™ extracellular matrix.

The cell lines used are HT 1080 (ATCC, Cat# CCL-121), DU-145 (ATCC, Cat#HTB-81), PC3 (ATCC, Cat# CRL-1435) or B16F1 (ATCC, Cat# CRL-6323).

The invasion test system is removed from the package from −20° C.storage and allowed to warm to ambient temperature. PBS is added to theinterior of the inserts and they are allowed to rehydrate for 2 hours at37° C. Then the medium is removed and 450 μL cell suspensions of tumourcells (grown to 50-70% confluence, trypsinized, and resuspended inmedium without serum at 1×10⁶/mL) is added to the top chamber. Testcompounds are added to the top chamber at 10× the desired finalconcentration in 50 μL volumes. DMSO acts as control.

Then 750 μL of medium containing 50% fresh growth medium with 10% FBSand 50% NIH 3T3-conditioned medium is added to each of the bottom wells.The invasion system is then incubated for 24 to 48 hours at 37° C., in a5% CO₂ atmosphere.

Following incubation, the insert plate is transferred into a second24-well plate containing 0.5 mL of 5 μg/mL calcein AM (Molecular Probes)in Hanks buffered salt solution (HBSS), and plates are incubated for 1hour at 37° C., 5% CO₂.

Fluorescence data indicating cell invasion is read in a FluoroskanAscent FL™ (LabSystems) with bottom reading at excitation/emissionwavelength of 485/538 nm.

Data is expressed as fluorescence unit (FU) from the sum of middle 25areas per 24-well or as percentage of invasion inhibition by followingformula: % of invasion inhibition−100−FU of compound treated cellinvasion/FU of DMSO treated cell invasion times 100.

The compounds inhibit invasion in this assay, and thus may be used toprevent metastasis in cancer and tissue remodeling.

EXAMPLE 9 Peritoneal Macrophage Stimulation and Analysis

A. Establishment of inflammation assay panel.

Macrophages are important elements of innate immunity to infection andare among the first cell type in the immune response to be exposed toand activated by infectious agents. IFN-γ and LPS are potent activatorsof macrophages, priming them for a variety of biological effects. IFN-γ,initially secreted by NK and T cells in response to infection, convertsmacrophages from a resting to an activated state (inflammatorymacrophages), priming them for antimicrobial activity manifested byincreased killing of intracellular pathogens, and antigen processing andpresentation to lymphocytes. The action of IFN-γ is synergized with theLPS second messenger, enhancing the stimulation of macrophages throughthe activation of NF-κB, that results in the transcriptionalup-regulation of a number of genes involved in the cell-mediated immuneresponse, including inducible iNOS (nitric oxide synthase). Activatedmacrophages are qualitatively different from quiescent macrophages.These differences are typically observed by an increased proliferationindex, up-regulated expression of MHC-II, and production of variousbioactive molecules. The latter biological effects are mediated by NO(nitric oxide) release and increased production of pro-inflammatorycytokines (IL-6, TNF-γ, IL-1). Primary macrophages derived from Balb/cmice and RAW 264.7 cells (Balb/c background) were used to establish invitro inflammatory models with fast and reliable readouts.

B. Materials and Methods

1. Reagents.

The iNOS inhibitor NG-monomethyl-L-arginine (L-NMMA) and murine rIFN-γare purchased from Calbiochem, (San Diego, Calif.). Protein-free,phenol/water-extracted LPS (from E. coli serotype 0111:B4 0127:B8),Zymosan A™, dexamethasone and hydrocortisone, sulfanilamide andN-(1-naphthyl)-ethylenediamine, arare purchased from Sigma (St. Louis,Mo.). Human recombinant vascular endothelial growth factor (VEGF) ispurchased from R&D Systems (Minneapolis, Minn.). Rabbit polyclonalantibody against active (phosphorylated) extracellular signal-regulatedkinase (ERK), as well as HRP-conjugated donkey anti-rabbit IgG areobtained from Promega (Madison, Wis.). ELISA dual-set kit for detectionof IL-6 is purchased from PharMingen (San Diego, Calif.). Anti-murineiNOS/NOS type II and cyclooxygenase-2 (COX-2) antibodies are obtainedfrom Transduction Laboratories (Lexington, Ky.).

Female BALB/c inbred mice, 6-12 weeks of age, are purchased from HarlanInc. (Indianapolis, Ind.) and housed under fluorescent light for 12 hper day. Mice are housed in cages, and maintained in compliance with theCanadian Council on Animal Care standards.

2. Isolation of Primary Mouse Macrophages.

Peritoneal exudate macrophages are isolated by peritoneal lavage withice-cold sterile physiological saline 24 hours after intraperitonealinjection of BALB/c mice with 0.5 mL of sterile Zymosan A™ (1 mg/0.5 mL0.9% saline). Cells are washed and resuspended in RPMI 1640 supplementedwith 1 mM D-glucose, 1 mM sodium pyruvate, 100 units/mL penicillin, 100μg/mL streptomycin, and 5% FBS.

3. Treatment of Primary Macrophages.

Primary macrophages (1.5×105 cells/well) are grown in 96-well plates(nitrite assay), or 6-well plates (2×106 cells/well) for measurement ofiNOS and COX-2 expression. Following 3 hours incubation, at 37° C., 5%CO₂ (allowing macrophages to attach) cells are stimulated with LPS (5μg/mL) and IFN-γ (100 U/mL) in the absence or presence of variousconcentrations of test compounds (all treatments are replicated sixtimes). Cells are incubated for an additional 24 hours, and cell freeculture supernatants from each well are collected for NO and cytokinedetermination. The remaining cells are stained with crystal violet orMTS to determine effect of the test compounds on cell survival.

4. NO Production.

Following stimulation, the production of NO is determined by assayingculture supernatants for NO₂, a stable reaction product of NO withmolecular oxygen. Briefly, 100 μL of culture supernatant is reacted withan equal volume of Griess reagent at ambient temperature for 10 minutes.The absorbance at 550 nm is determined. All measurements are performedsix times. The concentration of NO₂ is calculated based on comparisonwith a standard curve prepared using NaNO₂.

5. Western blot analysis.

After incubation with the indicated stimuli in the presence ofinhibitors, cells (duplicate samples, 2×10⁶ cell/6-wells plate) arewashed in PBS and lysed on ice in 60 μL of lysis buffer. The proteincontent of each sample is determined using the Bradford protein assaykit (Bio-Rad, Richmond, Calif.). Absorbance is measured at 750 nm with aBeckman DU530 spectrophotometer (Palo Alto, Calif.). Proteins are mixedwith 45×SDS sample buffer. Following separation of proteins by SDS-PAGE,using 8% bis-acrylamide in the separation gel, the proteins aretransferred from the gels onto PVDF membranes using a MiniProtean™ IIICell (Bio-Rad), at 100 V for 1.5 hours. Equal amounts of protein (5 μg)are loaded onto SDS-PAGE gels and examined by Western blot analysis withanti-actin, anti-iNOS, anti-COX-2 murine monoclonal antibodies,according to the manufacturer's specifications (TransductionLaboratories). Primary antibodies, in 5% blocking buffer (5% NFM/TTBS),are incubated with blots 2 hours or overnight at 4° C., followed byincubation with peroxidase-conjugated secondary antibody.Chemiluminescence substrates are used to reveal positive bands. Thebands are exposed on X-ray films. The films are used to analyze theimpact of inhibitors on expression of iNOS and COX-2 compared to variouscontrols and “house-keeping” protein (Actin) concentration to controlthe protein loading and detect any non-specific effects on proteinproduction. The Multi-Analyst™/PC system from Biorad is used toquantitate the bands of the expressed protein on the film. This versionof Multi-Analyst is used with the Bio-Rad Gel Doc 1000™ imaging system.White light is chosen as the selected light source, thus the signalstrength is measured in OD (optic density) units. The OD of each band isbeing subtracted to a global background area of the gel.

C. In Vitro Angiogenesis.

HUVEC cells cultured for 24 hours in M199 with 0.5% FCS are plated at6×105 cells/well in 12-well plates pre-coated with 300 μL of Matrigel(10.7 mg/mL; Becton Dickinson) in M199 with 0.5% FCS in the presence ofVEGF (1 ng/mL), and in the absence or presence of positive control(Z)-3-[2,4-dimethyl-5-(2-oxo-1,2-dihydroindol-3-ylidenemethyl)-1H-pyrrol-3-yl]propionicacid or various inhibitors. After 5 hours of incubation in a 5%CO₂-humidified atmosphere at 37° C., the three-dimensional organizationof the cells is examined using an inverted photomicroscope. The cellsare fixed with crystal violet (0.05% in 20% ethanol) and digitallyphotographed.

D. Enzyme Immunoassays for Mouse IL-6.

IL-6 levels are determined with PharMingen's OptEIA™ ELISA set developedusing an anti-mouse IL-6 Ab pair and mouse rIL-6 standard (PharMingen).Maxisorp F16™ multiwell strips (Nunc, Roskilde, Denmark) are coated withanti-mouse IL-6 capture Ab (at recommended concentration) in 0.1 MNaHCO₃, pH 9.5, 100 μL/well, overnight at 4° C. Plates are washed threetimes with 0.05% Tween 20 in PBS (PBST) and blocked for 1 hour atambient temperature with 200 μL/well of 10% FCS in PBS (blocking anddilution buffer). Plates are washed three times with PBST and duplicatesamples (100 μL/well) or standards (100 μL/well) in diluent buffer areincubated for 2 hours at ambient temperature. Plates are washed fivetimes with PBST and incubated with biotinylated anti-mouse IL-6 andavidin-horseradish peroxidase conjugate (at concentrations recommendedby the manufacturer) for 1 hour at ambient temperature. Plates arewashed seven times with PBST and 100 μL of 3,3′5,5′ tetramethylbenzidinesubstrate solution (TMB substrate reagent set, BD PharMingen) is addedto each well. After 15-30 minute incubation at ambient temperature,color development is terminated by adding 50 μL of 2N H₂SO₄ (Sigma).Absorbance is read at 450 nm with an EL 312e™ microplate reader or thelike. The lower limit of detection for IL-6 is 15.6 μg/mL.

EXAMPLE 10 NIDDM Model

In vivo oral treatment with pharmaceuticals compositions of theinvention result in significant glucose lowering in several rodentmodels of diabetes. In db/db mice, oral administration of the compoundselicited significant correction of hyperglycemia. In astreptozotocin-induced diabetic mouse model, compounds potentiate theglucose-lowering effect of insulin.

In normal rats, compounds improve oral glucose tolerance withsignificant reduction in insulin release following glucose challenge. Astructurally related inactive analog is not effective on insulinreceptor activation or glucose lowering in db/db mice.

1-23. (canceled)
 24. A pharmaceutical composition useful in treatingcancer, inflammation, diabetes or obesity associated with the activityof PTPN1 in a human, wherein the pharmaceutical composition comprises apharmaceutically acceptable carrier, diluent or excipient and a compoundof formula (Ia):

wherein: each p is independently 1 to 5; A is linker of four atoms andis selected from the group consisting of an optionally substitutedstraight or branched alkylene chain of four carbons, an optionallysubstituted straight or branched alkenylene chain of four carbons,—R^(8a)—C(R^(2a))—N(R^(5a))—, —R^(8a)—N(R^(5a))—C(R^(2a))—,R^(8a)—O—C(R^(2a))—, —R^(7a)—O—C(R^(2a))—R^(7a)—, —R^(8a)—O—R^(7a)—,—R^(7a)—C(R^(2a))—N(R^(5a))—S(O)_(t)— (where t is 0 to 2),—R^(8a)—N(R^(5a))—R^(7a), —R^(8a)—S(O)_(t)—R^(7a)— (where t is 0 to 2),—R^(9a)—N(R^(5a))—, —R^(9a)—O—, and R^(9a)—C(R^(2a))—; R^(1a) and R^(2a)are each independently ═O or ═S; R^(3a) is alkyl, alkenyl, aryl,aralkyl, aralkenyl, cycloalkyl, cycloalkylalkyl, chloro, iodo, bromo,haloalkyl, haloalkoxy, nitro, cyano, —N═N—O—R^(11a), —OR^(10a),—C(O)OR^(10a), —C(O)N(R^(10a))₂, —N(R^(12a))₂, —N(R^(10a))C(O)R^(10a),N(R^(10a))C(O)OR^(11a), —S(O)_(t)R^(10a) (where t is 0 to 2),—S(O)_(t)N(R^(10a))₂ (where t is 0 to 2), or heterocyclylalkyl; R^(4a)is hydrogen, alkyl, aralkyl, aryl, haloalkyl, cycloalkyl,cycloalkylalkyl, heterocyclyl or heterocyclylalkyl; R^(5a) is hydrogen,alkyl, aralkyl, or aryl; R^(6a) is alkyl, alkenyl, aryl, aralkyl,aralkenyl, cycloalkyl, cycloalkylalkyl, halo, haloalkyl, haloalkoxy,nitro, cyano, —N═N—O—R^(11a), C(O)N(R^(10a))₂, —N(R^(10a))₂,N(R^(10a))C(O)R^(10a), —N(R^(10a))C(O)OR^(11a), —S(O)_(t)R^(10a) (wheret is 0 to 2), —S(O)_(t)N(R^(10a))₂ (where t is 0 to 2),—S(O)_(t)NH—R^(14a), heterocyclyl or heterocyclylalkyl; each R^(7a) isan optionally substituted alkylene chain of one carbon; each R^(8a) isan optionally substituted straight or branched alkylene or alkenylenechain of two carbons; each R^(9a) is an optionally substituted alkylenechain of three carbons; each R^(10a) is hydrogen, alkyl, alkenyl,cycloalkyl, cycloalkylalkyl, aralkyl or aryl; R^(11a) is hydrogen, alkylor aralkyl; R¹² is hydrogen, aryl or aralkyl; and R^(14a) is a thiazole;as a single stereoisomer, a mixture of stereoisomers, or as a racemicmixture of stereoisomers; or as a solvate or polymorph; or as apharmaceutically acceptable salt thereof.
 25. The pharmaceuticalcomposition of claim 24 wherein the compound of formula (Ia) is acompound of formula (Ia) wherein: A is —R^(8a)—C(R^(2a))—N(R^(5a))—;R^(1a) and R^(2a) are each independently ═O or ═S; R^(3a) is alkyl,aryl, aralkyl, chloro, iodo, bromo, haloalkyl, haloalkoxy, nitro, cyano,—N═N—O—R^(11a), OR^(12a), —C(O)OR^(10a), —C(O)N(R^(10a))₂, —N(R^(12a))₂,—N(R^(10a))C(O)R^(10a), —N(R^(10a))C(O)OR^(11a), —S(O)_(t)R^(10a) (wheret is 0 to 2), or —S(O)_(t)N(R^(10a))₂ (where t is 0 to 2); R^(4a) ishydrogen or alkyl; R^(5a) is hydrogen or alkyl; R^(6a) is alkyl, aryl,aralkyl, halo, haloalkyl, haloalkoxy, nitro, cyano, —N═N—O—R^(11a),—C(O)N(R^(10a))₂, —N(R^(10a))₂, —N(R^(10a))C(O)R^(10a),—N(R^(10a))C(O)OR^(11a), —S(O)_(t)R^(10a) (where t is 0 to 2),—S(O)_(t)NH—R^(14a) or —S(O)_(t)N(R^(10a))₂ (where t is 0 to 2); eachR^(8a) is an optionally substituted straight or branched alkylene oralkenylene chain of two carbons; each R^(10a) is hydrogen, alkyl,aralkyl or aryl; R^(11a) is hydrogen, alkyl or aralkyl; R^(12a) ishydrogen, aryl or aralkyl; and R^(14a) is a thiazole.
 26. Thepharmaceutical composition of claim 25 wherein the compound of formula(Ia) is a compound of formula (Ia) wherein: A is—R^(8a)—C(R^(2a))—N(R^(5a))—; R^(1a) and R^(2a) are each independently═O or ═S; R³, is alkyl, aryl, aralkyl, chloro, iodo, bromo, haloalkyl,or haloalkoxy; R^(4a) is hydrogen or alkyl; R^(5a) is hydrogen or alkyl;R^(6a) is —C(O)N(R^(10a))₂, —S(O)_(t)R^(10a) (where t is 0 to 2),—S(O)_(t)NH—R^(14a) or —S(O)_(t)N(R^(10a))₂ (where t is 0 to 2); eachR^(8a) is an optionally substituted straight or branched alkylene oralkenylene chain of two carbons; R^(14a) is a thiazole; and each R^(10a)is hydrogen, alkyl, aralkyl or aryl.
 27. The pharmaceutical compositionof claim 26 wherein the compound of formula (Ia) is a compound offormula (Ia) wherein: A is —R^(8a)—C(R^(2a))—N(R^(5a))—; R^(1a) andR^(2a) are both ═O; R^(3a) is alkyl, haloalkyl or haloalkoxy; R^(4a) ishydrogen; R^(5a) is hydrogen or alkyl; R^(6a) is —S(O)₂N(R¹⁰)₂; R^(8a)is ethylene; and each R^(10a) is hydrogen or alkyl.
 28. A method oftreating cancer in a mammal, which method comprises administering to themammal in need thereof a therapeutically effective amount of a compoundof formula (I):

wherein: each p is independently 1 to 5; A is a linker of four atoms andis selected from the group consisting of an optionally substitutedstraight or branched alkylene chain of four carbons, an optionallysubstituted straight or branched alkenylene chain of four carbons,—R⁸—C(R²)—N(R⁵)—, —R⁸—N(R⁵)—C(R²)—, —R⁸—O—C(R²)—, —R⁷—O—C(R²)—R⁷—,—R⁸—O—R⁷—, —R⁷—C(R²)—N(R⁵)—S(O)_(t)— (where t is 0 to 2), —R⁸—N(R⁵)—R⁷,—R⁸—S(O)_(t)—R⁷— (where t is 0 to 2), —R⁹—N(R⁵)—, —R⁹—O—, and—R⁹—C(R²)—; R¹ and R² are each independently ═O or ═S; each R³ and R⁶are independently selected from the group consisting of hydrogen, alkyl,alkenyl, aryl, aralkyl, aralkenyl, cycloalkyl, cycloalkylalkyl, halo,haloalkyl, haloalkoxy, nitro, cyano, —N═N—O—R¹¹, —OR¹⁰, —C(O)OR¹⁰,—C(O)N(R¹⁰)₂, —N(R¹⁰)₂, —N(R¹⁰)C(O)R¹⁰, —N(R¹⁰)C(O)OR¹¹, —S(O)_(t)R¹⁰(where t is 0 to 2), —S(O)_(t)N(R¹⁰)₂ (where t is 0 to 2),—S(O)_(t)NH—R¹⁴, heterocyclyl and heterocyclylalkyl; R⁴ is hydrogen,alkyl, aralkyl, aryl, haloalkyl, cycloalkyl, cycloalkylalkyl,heterocyclyl or heterocyclylalkyl; each R⁵ is independently hydrogen,alkyl, aralkyl, or aryl; each R⁷ is an optionally substituted alkylenechain of one carbon; each R⁸ is an optionally substituted straight orbranched alkylene or alkenylene chain of two carbons; each R⁹ is anoptionally substituted alkylene chain of three carbons; each R¹⁰ ishydrogen, alkyl, alkenyl, cycloalkyl, cycloalkylalkyl, aralkyl or aryl;R¹¹ is hydrogen, alkyl or aralkyl; and R¹⁴ is a thiazole; as a singlestereoisomer, a mixture of stereoisomers, or as a racemic mixture ofstereoisomers; or as a solvate or polymorph; or as a pharmaceuticallyacceptable salt thereof.
 29. The method of claim 28 wherein the mammalis a human.
 30. The method of claim 29 wherein the cancer is associatedwith hyperproliferation or tissue remodelling or repair.
 31. The methodof claim 30 wherein the cancer is associated with the activity of PTPN12or PTPN2.
 32. The method of any one of claims 28-31 wherein the compoundof formula (I) is a compound of formula (I) wherein: A is—R⁸—C(R²)—N(R⁵)—; R¹ and R² are each independently ═O or ═S; R³ and R⁶are each independently selected from the group consisting of hydrogen,alkyl, aryl, aralkyl, halo, haloalkyl, haloalkoxy, nitro, cyano,—N═N—O—R”—, —OR¹⁰, —C(O)OR¹⁰, —C(O)N(R¹⁰)₂, —N(R¹⁰)₂, —N(R¹⁰)C(O)R¹⁰,—N(R¹⁰)C(O)OR¹¹, —S(O)_(t)R¹⁰ (where t is 0 to 2), —S(O)_(t)N(R¹⁰)₂(where t is 0 to 2), —S(O)_(t)NH—R¹⁴, and R⁴ is hydrogen or alkyl; R⁵ ishydrogen or alkyl; R⁸ is an optionally substituted straight or branchedalkylene or alkenylene chain of two carbons; each R¹⁰ is hydrogen,alkyl, aralkyl or aryl; R¹¹ is hydrogen, alkyl or aralkyl; and R¹⁴ is athiazole.
 33. The method of claim 32 wherein the compound of formula (I)is a compound of formula (I) wherein: A is —R⁸—C(R²)—N(R⁵)—; R¹ and R²are each independently ═O or ═S; R³ is hydrogen, alkyl, alkoxy, aryl,aralkyl, halo, haloalkyl, or haloalkoxy; R⁴ is hydrogen or alkyl; R⁵ ishydrogen or alkyl; R⁶ is —C(O)OR¹⁰, —C(O)N(R¹⁰)₂, —S(O)_(t)R¹⁰ (where tis 0 to 2), —S(O)_(t)N(R¹⁰)₂ (where t is 0 to 2), or —S(O)_(t)NH—R¹⁴; R⁸is an optionally substituted straight or branched alkylene or alkenylenechain of two carbons; each R¹⁰ is hydrogen, alkyl, aralkyl or aryl; andR¹⁴ is a thiazole.
 34. The method of claim 33 wherein the compound offormula (I) is a compound of formula (I) wherein: A is —R⁸—C(R²)—N(R⁵);R¹ and R² are both ═O; R³ is alkyl, alkoxy, haloalkyl or haloalkoxy; R⁴is hydrogen; R⁵ is hydrogen or alkyl; R⁶ is —S(O)₂N(R¹⁰)₂; R⁸ isethylene; and R¹⁰ is hydrogen or alkyl.
 35. A method of treatinginflammation in a mammal, which method comprises administering to themammal in need thereof a therapeutically effective amount of a compoundof formula (Ia):

wherein: each p is independently 1 to 5; A is linker of four atoms andis selected from the group consisting of an optionally substitutedstraight or branched alkylene chain of four carbons, an optionallysubstituted straight or branched alkenylene chain of four carbons,—R^(8a)—C(R^(2a))—N(R^(5a))—, —R^(8a)—N(R^(5a))—C(R^(2a))—,—R^(8a)—O—C(R^(2a)), —R^(7a)—O—C(R^(2a))—R^(7a)—, —R^(8a)—O—R^(7a),—R^(7a)—C(R^(2a))—N(R^(5a))—S(O)_(t)— (where t is 0 to 2),—R^(8a)—N(R^(5a))—R^(7a), —R^(8a)—S(O)_(t)—R^(7a)— (where t is 0 to 2),—R^(9a)—N(R^(5a))—, —R^(9a)—O—, and R^(9a)—C(R^(2a))—; R^(1a) and R^(2a)are each independently ═O or ═S; R^(3a) is alkyl, alkenyl, aryl,aralkyl, aralkenyl, cycloalkyl, cycloalkylalkyl, chloro, iodo, bromo,haloalkyl, haloalkoxy, nitro, cyano, —N═N—O—R^(11a), —OR^(10a),—C(O)OR^(10a), —C(O)N(R^(10a))₂, —N(R^(12a))₂, —N(R^(10a))C(O)R^(10a),—N(R^(10a))C(O)OR^(11a), S(O)_(t)R^(10a) (where t is 0 to 2),—S(O)_(t)N(R^(10a))₂ (where t is 0 to 2), or heterocyclylalkyl; R^(4a)is hydrogen, alkyl, aralkyl, aryl, haloalkyl, cycloalkyl,cycloalkylalkyl, heterocyclyl or heterocyclylalkyl; R^(5a) is hydrogen,alkyl, aralkyl, or aryl; R^(6a) is alkyl, alkenyl, aryl, aralkyl,aralkenyl, cycloalkyl, cycloalkylalkyl, halo, haloalkyl, haloalkoxy,nitro, cyano, —N═N—O—R^(11a), —C(O)N(R^(10a))₂, —N(R^(10a))₂,—N(R^(10a))C(O)R^(10a), —N(R^(10a))C(O)OR^(11a), —S(O)_(t)R^(10a) (wheret is 0 to 2), —S(O)_(t)N(R^(10a))₂ (where t is 0 to 2),—S(O)_(t)NH—R^(14a), heterocyclyl or heterocyclylalkyl; each R^(7a) isan optionally substituted alkylene chain of one carbon; each R^(8a) isan optionally substituted straight or branched alkylene or alkenylenechain of two carbons; each R^(9a) is an optionally substituted alkylenechain of three carbons; each R^(10a) is hydrogen, alkyl, alkenyl,cycloalkyl, cycloalkylalkyl, aralkyl or aryl; R^(11a) is hydrogen, alkylor aralkyl; R^(12a) is hydrogen, aryl or aralkyl; and R^(14a) is athiazole; as a single stereoisomer, a mixture of stereoisomers, or as aracemic mixture of stereoisomers; or as a solvate or polymorph; or as apharmaceutically acceptable salt thereof.
 36. The method of claim 35wherein the mammal is a human.
 37. The method of claim 36 wherein theinflammation is associated with hyperproliferation or tissue remodellingor repair.
 38. The method of claim 37 wherein the inflammation isassociated with the activity of PTPN12 or PTPN2.
 39. The method ofclaims 35-38 wherein the compound of formula (Ia) is a compound offormula (Ia) wherein: A is —R^(8a)—C(R^(2a))—N(R^(5a))—; R^(1a) andR^(2a) are each independently ═O or ═S; R^(3a) is alkyl, aryl, aralkyl,chloro, iodo, bromo, haloalkyl, haloalkoxy, nitro, cyano,—N═N—O—R^(11a), OR^(12a) —C(O)OR^(10a), —C(O)N(R^(10a))₂,—N(R^(12a))—N(R^(10a))C(O)R^(10a)—N(R^(10a))C(O)OR^(11a),—S(O)_(t)R^(10a) (where t is 0 to 2), or —S(O)_(t)N(R^(10a))₂ (where tis 0 to 2); R^(4a) is hydrogen or alkyl; R^(5a) is hydrogen or alkyl;R^(6a) is alkyl, aryl, aralkyl, halo, haloalkyl, haloalkoxy, nitro,cyano, —N═N—O—R^(1a), C(O)N(R^(10a))₂, —N(R^(10a))₂,—N(R^(10a))C(O)R^(10a), —N(R^(10a))C(O)OR^(11a), —S(O)_(t)R^(10a) (wheret is 0 to 2), —S(O)_(t)NH—R^(14a), or —S(O)_(t)N(R^(10a))₂ (where t is 0to 2); each R^(8a) is an optionally substituted straight or branchedalkylene or alkenylene chain of two carbons; each R^(10a) is hydrogen,alkyl, aralkyl or aryl; R^(11a) is hydrogen, alkyl or aralkyl; R^(12a)is hydrogen, aryl or aralkyl; and R^(14a) is a thiazole.
 40. The methodof claim 39 wherein the compound of formula (Ia) is a compound offormula (Ia) wherein: A is —R^(8a)—C(R^(2a))—N(R^(5a))—; R^(1a) andR^(2a) are each independently ═O or ═S; R^(3a) is alkyl, aryl, aralkyl,chloro, iodo, bromo, haloalkyl, or haloalkoxy; R^(4a) is hydrogen oralkyl; R^(5a) is hydrogen or alkyl; R^(6a) is —C(O)N(R^(10a))₂,—S(O)_(t)R^(10a) (where t is 0 to 2), —S(O)_(t)NH—R^(14a), or—S(O)_(t)N(R^(10a))₂ (where t is 0 to 2); each R^(8a) is an optionallysubstituted straight or branched alkylene or alkenylene chain of twocarbons; each R^(10a) is hydrogen, alkyl, aralkyl or aryl; and R^(14a)is a thiazole.
 41. The method of claim 40 wherein the compound offormula (Ia) is a compound of formula (Ia) wherein: A is—R^(8a)—C(R^(2a))—N(R^(5a))—; R^(1a) and R^(2a) are both ═O; R^(3a) isalkyl, haloalkyl or haloalkoxy; R^(4a) is hydrogen; R^(5a) is hydrogenor alkyl; R^(6a) is —S(O)₂N(R¹⁰)₂; R^(8a) is ethylene; and each R^(10a)is hydrogen or alkyl.
 42. A method of treating a mammal having adisorder or condition associated with hyperproliferation and tissueremodelling or repair, wherein said method comprises administering tothe mammal having the disorder or condition a therapeutically effectiveamount of a compound of formula (I):

wherein: each p is independently 1 to 5; A is a linker of four atoms andis selected from the group consisting of an optionally substitutedstraight or branched alkylene chain of four carbons, an optionallysubstituted straight or branched alkenylene chain of four carbons,—R⁸—C(R²)—N(R⁵)—, —R⁸—N(R⁵)—C(R²)—, —R⁸—O—C(R²)—, —R⁷—O—C(R²)—R⁷—,—R⁸—O—R⁷—, —R⁷—C(R²)—N(R⁵)—S(O)_(t)— (where t is 0 to 2), —R⁸—N(R⁵)—R⁷,—R⁸—S(O)_(t)—R⁷— (where t is 0 to 2), —R⁹—N(R⁵)—, —R⁹—O—, and—R⁹—C(R²)—; R¹ and R² are each independently ═O or ═S; each R³ and R⁶are independently selected from the group consisting of hydrogen, alkyl,alkenyl, aryl, aralkyl, aralkenyl, cycloalkyl, cycloalkylalkyl, halo,haloalkyl, haloalkoxy, nitro, cyano, —N═N—O—R¹¹, —OR¹⁰, —C(O)OR¹⁰,—C(O)N(R¹⁰)₂, —N(R¹⁰)₂, —N(R¹⁰)C(O)R¹⁰, —N(R¹⁰)C(O)OR¹¹, —S(O)_(t)R¹⁰(where t is 0 to 2), —S(O)_(t)N(R¹⁰)₂ (where t is 0 to 2),—S(O)_(t)NH—R¹⁴, heterocyclyl and heterocyclylalkyl; R⁴ is hydrogen,alkyl, aralkyl, aryl, haloalkyl, cycloalkyl, cycloalkylalkyl,heterocyclyl or heterocyclylalkyl; each R⁵ is independently hydrogen,alkyl, aralkyl, or aryl; each R⁷ is an optionally substituted alkylenechain of one carbon; each R⁸ is an optionally substituted straight orbranched alkylene or alkenylene chain of two carbons; each R⁹ is anoptionally substituted alkylene chain of three carbons; each R¹⁰ ishydrogen, alkyl, alkenyl, cycloalkyl, cycloalkylalkyl, aralkyl or aryl;R¹¹ is hydrogen, alkyl or aralkyl; and R¹⁴ is a thiazole; as a singlestereoisomer, a mixture of stereoisomers, or as a racemic mixture ofstereoisomers; or as a solvate or polymorph; or as a pharmaceuticallyacceptable salt thereof.
 43. A method of treating a mammalian cell witha compound of formula (I):

wherein: each p is independently 1 to 5; A is a linker of four atoms andis selected from the group consisting of an optionally substitutedstraight or branched alkylene chain of four carbons, an optionallysubstituted straight or branched alkenylene chain of four carbons,—R⁸—C(R²)—N(R⁵)—, —R⁸—N(R⁵)—C(R²)—, —R⁸—O—C(R²)—, —R⁷—O—C(R²)—R⁷—,—R⁸—O—R⁷—, —R⁷—C(R²)—N(R⁵)—S(O)_(t)— (where t is 0 to 2), —R⁸—N(R⁵)—R⁷,—R⁸—S(O)_(t)—R⁷— (where t is 0 to 2), —R⁹—N(R⁵)—, —R⁹—O—, and—R⁹—C(R²)—; R¹ and R² are each independently ═O or ═S; each R³ and R⁶are independently selected from the group consisting of hydrogen, alkyl,alkenyl, aryl, aralkyl, aralkenyl, cycloalkyl, cycloalkylalkyl, halo,haloalkyl, haloalkoxy, nitro, cyano, —N═N—O—R¹¹, —OR¹¹, —C(O)OR¹⁰,—C(O)N(R¹⁰)₂, —N(R¹⁰)₂, —N(R¹⁰)C(O)R¹⁰, —N(R¹⁰)C(O)OR¹¹, —S(O)_(t)R¹⁰(where t is 0 to 2), —S(O)_(t)N(R¹⁰)₂ (where t is 0 to 2),—S(O)_(t)NH—R¹⁴, heterocyclyl and heterocyclylalkyl; R⁴ is hydrogen,alkyl, aralkyl, aryl, haloalkyl, cycloalkyl, cycloalkylalkyl,heterocyclyl or heterocyclylalkyl; each R⁵ is independently hydrogen,alkyl, aralkyl, or aryl; each R⁷ is an optionally substituted alkylenechain of one carbon; each R⁸ is an optionally substituted straight orbranched alkylene or alkenylene chain of two carbons; each R⁹ is anoptionally substituted alkylene chain of three carbons; each R¹⁰ ishydrogen, alkyl, alkenyl, cycloalkyl, cycloalkylalkyl, aralkyl or aryl;R¹¹ is hydrogen, alkyl or aralkyl; and R¹⁴ is a thiazole; as a singlestereoisomer, a mixture of stereoisomers, or as a racemic mixture ofstereoisomers; or as a solvate or polymorph; or as a pharmaceuticallyacceptable salt thereof; wherein the method comprises administering thecompound of formula (I) to a mammalian cell and the compound of formula(I) is capable of inhibiting the activity of PTPN1, PTPN12 or PTPN2within the mammalian cell.
 44. The method of claim 43 wherein themammalian cell is treated in vitro.
 45. The method of claim 44 whereinthe mammalian cell is treated in vivo.
 46. The method of claim 45wherein the inhibition of activity results in a reduction of celladhesion.
 47. The method of claim 45 wherein the inhibition of activityresults in a reduction of cell division.
 48. The method of claim 45,wherein the inhibition of activity results in a reduction of cellmigration.
 49. The method of claim 45, wherein the inhibition ofactivity results in control of tumor growth.
 50. The method of claim 45wherein the inhibition of activity results in control of lymphocyteactivation.
 51. A method of treating diabetes in a mammal, which methodcomprises administering to the mammal in need thereof a therapeuticallyeffective amount of a compound of formula (I):

wherein: each p is independently 1 to 5; A is a linker of four atoms andis selected from the group consisting of an optionally substitutedstraight or branched alkylene chain of four carbons, an optionallysubstituted straight or branched alkenylene chain of four carbons,—R⁸—C(R²)—N(R⁵)—, —R⁸—N(R⁵)—C(R²)—, —R⁸—O—C(R²)—, —R⁷—O—C(R²)—R⁷—,—R⁸—O—R⁷—, —R⁷—C(R²)—N(R⁵)—S(O)_(t)— (where t is 0 to 2), —R⁸—N(R⁵)—R⁷,—R⁸—S(O)_(t)—R⁷— (where t is 0 to 2), —R⁹—N(R⁵)—, —R⁹—O—, and—R⁹—C(R²)—; R¹ and R² are each independently ═O or ═S; each R³ and R⁶are independently selected from the group consisting of hydrogen, alkyl,alkenyl, aryl, aralkyl, aralkenyl, cycloalkyl, cycloalkylalkyl, halo,haloalkyl, haloalkoxy, nitro, cyano, —N═N—O—R¹¹, —OR¹⁰, —C(O)OR¹⁰,—C(O)N(R¹⁰)₂, —N(R¹⁰)₂, —N(R¹⁰)C(O)R¹⁰, —N(R¹⁰)C(O)OR¹¹, —S(O)_(t)R¹⁰(where t is 0 to 2), —S(O)_(t)N(R¹⁰)₂ (where t is 0 to 2),—S(O)_(t)NH—R¹⁴, heterocyclyl and heterocyclylalkyl; R⁴ is hydrogen,alkyl, aralkyl, aryl, haloalkyl, cycloalkyl, cycloalkylalkyl,heterocyclyl or heterocyclylalkyl; each R⁵ is independently hydrogen,alkyl, aralkyl, or aryl; each R⁷ is an optionally substituted alkylenechain of one carbon; each R⁸ is an optionally substituted straight orbranched alkylene or alkenylene chain of two carbons; each R⁹ is anoptionally substituted alkylene chain of three carbons; each R¹⁰ ishydrogen, alkyl, alkenyl, cycloalkyl, cycloalkylalkyl, aralkyl or aryl;R¹¹ is hydrogen, alkyl or aralkyl; and R¹⁴ is a thiazole; as a singlestereoisomer, a mixture of stereoisomers, or as a racemic mixture ofstereoisomers; or as a solvate or polymorph; or as a pharmaceuticallyacceptable salt thereof.
 52. The method of claim 51 wherein the mammalis a human.
 53. The method of claim 51 wherein the diabetes isassociated with the activity of PTPN1.
 54. The method of any one ofclaims 51-53 wherein the compound of formula (I) is a compound offormula (I) wherein: A is —R⁸—C(R²)—N(R⁵); R¹ and R² are eachindependently ═O or ═S; R³ and R⁶ are each independently selected fromthe group consisting of hydrogen, alkyl, aryl, aralkyl, halo, haloalkyl,haloalkoxy, nitro, cyano, —N═N—O—R¹¹—, —OR¹⁰, —C(O)OR¹⁰, —C(O)N(R¹⁰)₂,—N(R¹⁰)₂, —N(R¹⁰)C(O)R¹⁰, —N(R¹⁰)C(O)OR¹¹, —S(O)_(t)R¹⁰ (where t is 0 to2), —S(O)_(t)N(R¹⁰)₂ (where t is 0 to 2), —S(O)_(t)NH—R¹⁴, and R⁴ ishydrogen or alkyl; R⁵ is hydrogen or alkyl; R⁸ is an optionallysubstituted straight or branched alkylene or alkenylene chain of twocarbons; each R¹⁰ is hydrogen, alkyl, aralkyl or aryl; R¹¹ is hydrogen,alkyl or aralkyl; and R¹⁴ is a thiazole.
 55. The method of claim 54wherein the compound of formula (I) is a compound of formula (I)wherein: A is —R⁸—C(R²)—N(R⁵)—; R¹ and R² are each independently ═O or═S; R³ is hydrogen, alkyl, alkoxy, aryl, aralkyl, halo, haloalkyl, orhaloalkoxy; R⁴ is hydrogen or alkyl; R⁵ is hydrogen or alkyl; R⁶ is—C(O)OR¹⁰, —C(O)N(R¹⁰)₂, —S(O)_(t)R¹⁰ (where t is 0 to 2),—S(O)_(t)N(R¹⁰)₂ (where t is 0 to 2), or —S(O)_(t)NH—R¹⁴; R⁸ is anoptionally substituted straight or branched alkylene or alkenylene chainof two carbons; each R¹⁰ is hydrogen, alkyl, aralkyl or aryl; and R¹⁴ isa thiazole.
 56. The method of claim 55 wherein the compound of formula(I) is a compound of formula (I) wherein: A is —R⁸—C(R²)—N(R⁵)—; R¹ andR² are both ═O; R³ is alkyl, alkoxy, haloalkyl or haloalkoxy; R⁴ ishydrogen; R⁵ is hydrogen or alkyl; R⁶ is —S(O)₂N(R¹⁰)₂; R⁸ is ethylene;and R¹⁰ is hydrogen or alkyl.
 57. A method of treating obesityassociated with the activity of PTPN1 in a mammal, which methodcomprises administering to the mammal in need thereof a therapeuticallyeffective amount of a compound of formula (Ia):

wherein: each p is independently 1 to 5; A is linker of four atoms andis selected from the group consisting of an optionally substitutedstraight or branched alkylene chain of four carbons, an optionallysubstituted straight or branched alkenylene chain of four carbons,—R^(8a)—C(R^(2a))—N(R^(5a))—, —R^(8a)-N(R^(5a))—C(R^(2a))—,R^(8a)—O—C(R^(2a))—, R^(7a)—O—C(R^(2a))—R^(7a)—, —R^(8a)-O-R^(7a)—,—R^(7a)—C(R^(2a))—N(R^(5a))—S(O)_(t)— (where t is 0 to 2),—R^(8a)—N(R^(5a))—R^(7a), —R^(8a)—S(O)_(t)—R^(7a)— (where t is 0 to 2),—R^(9a)—N(R^(5a))—, and —R^(9a)—C(R^(2a))—; R^(1a) and R^(2a) are eachindependently ═O or ═S; R^(3a) is alkyl, alkenyl, aryl, aralkyl,aralkenyl, cycloalkyl, cycloalkylalkyl, chloro, iodo, bromo, haloalkyl,haloalkoxy, nitro, cyano, —N═N—O—R^(11a), —OR^(10a), —C(O)OR^(10a),C(O)N(R^(10a))₂, —N(R^(12a))₂, —N(R^(10a))C(O)R^(10a),—N(R^(10a))C(O)OR^(11a), —S(O)_(t)R^(10a) (where t is 0 to 2),—S(O)_(t)N(R^(10a))₂ (where t is 0 to 2), or heterocyclylalkyl; R^(4a)is hydrogen, alkyl, aralkyl, aryl, haloalkyl, cycloalkyl,cycloalkylalkyl, heterocyclyl or heterocyclylalkyl; R^(5a) is hydrogen,alkyl, aralkyl, or aryl; R^(6a) is alkyl, alkenyl, aryl, aralkyl,aralkenyl, cycloalkyl, cycloalkylalkyl, halo, haloalkyl, haloalkoxy,nitro, cyano, —N═N—O—R^(11a), —C(O)N(R^(10a))₂, —N(R^(10a))₂,—N(R^(10a))C(O)R^(10a), —N(R^(10a))C(O)OR^(11a), —S(O)_(t)R^(10a) (wheret is 0 to 2), —S(O)_(t)N(R^(10a))₂ (where t is 0 to 2),—S(O)_(t)NH—R^(14a), heterocyclyl or heterocyclylalkyl; each R^(7a) isan optionally substituted alkylene chain of one carbon; each R^(8a) isan optionally substituted straight or branched alkylene or alkenylenechain of two carbons; each R^(9a) is an optionally substituted alkylenechain of three carbons; each R^(10a) is hydrogen, alkyl, alkenyl,cycloalkyl, cycloalkylalkyl, aralkyl or aryl; R^(11a) is hydrogen, alkylor aralkyl; R^(12a) is hydrogen, aryl or aralkyl; and R^(14a) is athiazole; as a single stereoisomer, a mixture of stereoisomers, or as aracemic mixture of stereoisomers; or as a solvate or polymorph; or as apharmaceutically acceptable salt thereof.
 58. The method of claim 57wherein the mammal is a human.
 59. The method of claim 57 wherein thecompound of formula (Ia) is a compound of formula (Ia) wherein: A is—R^(8a)—C(R^(2a))—N(R^(5a))—; R^(1a) and R^(2a) are each independently═O or ═S; R^(3a) is alkyl, aryl, aralkyl, chloro, iodo, bromo,haloalkyl, haloalkoxy, nitro, cyano, —N═N—O—R^(11a), —OR^(12a),—C(O)OR^(10a), —C(O)N(R^(10a))₂, —N(R^(12a))₂, —N(R^(10a))C(O)R^(10a),—N(R^(10a))C(O)OR^(11a), —S(O)_(t)R^(10a) (where t is 0 to 2), or—S(O)_(t)N(R^(10a))₂ (where t is 0 to 2); R^(4a) is hydrogen or alkyl;R^(5a) is hydrogen or alkyl; R^(6a) is alkyl, aryl, aralkyl, halo,haloalkyl, haloalkoxy, nitro, cyano, —N═N—O—R^(11a), —C(O)N(R^(10a))₂,—N(R^(10a))₂, —N(R^(10a))C(O)R^(10a), —N(R^(10a))C(O)OR^(11a),—S(O)_(t)R^(10a) (where t is 0 to 2), S(O)_(t)NH—R⁴², or—S(O)_(t)N(R^(10a))₂ (where t is 0 to 2); each R^(8a) is an optionallysubstituted straight or branched alkylene or alkenylene chain of twocarbons; each R^(10a) is hydrogen, alkyl, aralkyl or aryl; R^(11a) ishydrogen, alkyl or aralkyl; R^(12a) is hydrogen, aryl or aralkyl; andR^(14a) is a thiazole.
 60. The method of claim 59 wherein the compoundof formula (Ia) is a compound of formula (Ia) wherein: A is—R^(8a)—C(R^(2a))—N(R^(5a))—; R^(1a) and R^(2a) are each independently═O or ═S; R^(3a) is alkyl, aryl, aralkyl, chloro, iodo, bromo,haloalkyl, or haloalkoxy; R^(4a) is hydrogen or alkyl; R^(5a) ishydrogen or alkyl; R^(6a) is —C(O)N(R^(10a))₂, —S(O)_(t)R^(10a) (where tis 0 to 2), —S(O)_(t)NH—R^(14a), or —S(O)_(t)N(R^(10a))₂ (where t is 0to 2); each R^(8a) is an optionally substituted straight or branchedalkylene or alkenylene chain of two carbons; each R^(10a) is hydrogen,alkyl, aralkyl or aryl; and R^(14a) is a thiazole.
 61. The method ofclaim 60 wherein the compound of formula (Ia) is a compound of formula(Ia) wherein: A is —R^(8a)—C(R^(2a))—N(R^(5a))—; R^(1a) and R^(2a) areboth ═O; R^(3a) is alkyl, haloalkyl or haloalkoxy; R^(4a) is hydrogen;R^(5a) is hydrogen or alkyl; R^(6a) is —S(O)₂N(R^(10a))₂; R^(8a) isethylene; and each R^(10a) is hydrogen or alkyl.